C8-C14 Flashcards

1
Q

extracellular form of a virus

A

virion

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2
Q

protein shell of the virus

A

capsid

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3
Q

The virus redirects the host cell’s metabolism from growth to support virus replication and the assembly of new virions

A

virulent/ lytic infection

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4
Q

The host cell is not destroyed but is genetically altered
because the viral genome becomes part of the host genome.

A

lysogenic infection

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5
Q

Viral genomes of this configuration have the exact same base sequence as that of the viral mRNA that will be translated to form viral proteins.

A

plus configuration

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6
Q

Viral genomes of this configuration are complementary in base sequence to viral mRNA.

A

minus config

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7
Q

symmetry of rod-shaped viruses

A

helical symmetry

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8
Q

symmetry of spherical viruses

A

icosahedral symmetry

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9
Q

nucleic acid polymerases carried by RNA viruses that function to replicate the viral RNA genome and produce viral-specific mRNA

A

RNA dependent RNA polymerases- RNA replicases

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9
Q

example of enzyme carried by a virus that aid in their infection on the host cell

A

influenza virushas envelope proteins called neuraminidases that destroy glycoproteins and glycolipids of animal cell connective tissue, thus liberating the virions

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10
Q

carried by retroviral virions to make DNA from an RNA template

A

RdDp reverse transcriptase

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11
Q

viral life cycle

A
  1. Attachment (adsorption) of the virion to the host cell
  2. Penetration (entry, injection) of the virion nucleic acid into the
    host cell
  3. Synthesis of virus nucleic acid and protein by host cell machinery as redirected by the virus
  4. Assembly of capsids and packaging of viral genomes into new
    virions
  5. Release of new virions from the cell
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12
Q

number of virions release per cell

A

burst size

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13
Q

the number of infectious virions present per volume of fluid

A

titer

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14
Q

When a virus infects host cells growing on a flat surface, it creates a zone of cell lysis

A

plaque

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15
Q

efficiency of which virions infect host cells is rarely 100% and often considerably less. state reasons why

A

Virions that fail to infect may have assembled incompletely during the maturation process,
may contain defective genomes,
or may have suffered a spontaneous mutation that prevents them from attaching or otherwise properly replicating

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16
Q

example mechanisms of Archaea and bacteria against viral attack

A

toxin-antitoxin module
CRISPR
destruction of dsDNA through restriction endonucleases (process called restriction)

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17
Q

Besides encoding its own replication machinery, the T4 genome has another unusual feature: In a population of T4 virions, although each copy of the genome contains the same set of genes, they are arranged in a different order.

A

circular permutation

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18
Q

The T4 genome encodes three major
sets of proteins

A

early proteins, middle proteins, and late proteins

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19
Q

example of a virulent virus

A

Bacteriophage T4

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20
Q

some double-stranded DNA bacte rial viruses, although capable of a virulent cycle, can also infect
their host and establish a long-term stable relationship

A

temperate viruses

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21
Q

examples of temperate phages

A

lambda and P1

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22
Q

Maintenance of the lysogenic state is due to

A

phage encoded repressor protein

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23
Q

If lambda enters the lytic pathway, long, linear concatemers of genomic DNA are synthesized by a mechanism called

A

rolling circle replication

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24
Instead of the lytic pathway, if lambda takes the lysogenic route, its genome integrates into the E. coli chromosome requiring a protein called
lambda integrase
25
Whether lysis or lysogeny occurs in a lambda infection depends in large part on the levels of two key repressor proteins that can accumulate in the cell following infection which are
lambda repressor, also called the cI protein, and a second repressor called Cro
26
it represses the transcription of all other lambda-encoded genes, including Cro. When this happens, the lambda genome integrates into the host’s genome and becomes a prophage
cI protein
27
represses expression of a protein called cII whose function is to activate the synthesis of cI. Hence, following infection, if cI is present at insufficient levels to repress expression of phage-specific genes, this protein can accumulate in the cell; if this happens, lambda travels the lytic pathway
Cro
28
If an animal virus initially evades the immune system, animal viruses can catalyze at least four different outcomes
-A virulent infection results in lysis of the host cell; this is the most common outcome. -a latent infection, the viral DNA exists in the host’s genome and virions are not produced; this leaves the host cell unharmed unless and until an event triggers the virulent pathway. -With some enveloped animal viruses, release of virions, which occurs by a kind of budding process, may be slow, and the host cell may not be lysed (and thus not killed); instead it continues to grow and produce more virions. Such infections are called persistent infections. -Finally, certain animal viruses can convert a normal cell into a tumor cell, a process called transformation
29
structurally complex animal viruses that con tain an RNA genome. However, unlike other RNA viruses, the genome is replicated inside the host cell by way of a DNA intermediate
retroviruses
30
Retroviruses are enveloped viruses and carry several enzymes within the virion including
reverse transcriptase, integrase, and retroviral specific protease
31
the primary viruses infecting prokaryotic cells
Double-stranded DNA viruses (class I)
32
the major viral predators of eukaryotic cells
single-stranded plus-sense RNA viruses (class IV)
33
fungi are only infected by what class of viruses
Class III and IV
34
the vast majority of class I viruses that infect eukaryotes replicate in what hosts
Animal hosts rather than plants
35
hosts of class II viruses
more plants rather than animals
36
host of class V
animals rather than plants
37
host of class VI
known only from animal hosts
38
class VII host
much more common in plants than in animals
39
virus infecting the protozoan Acanthamoeba and belongs to a group of giant viruses with large genomes called nucleocytoplasmic large DNA viruses (NCLDV); multilayered and icosahedral capsid
mimivirus
40
comprise several virus families, including pox viruses, iridoviruses, and certain plant viruses. These viruses share a set of highly homologous proteins, most of which function in DNA metabolism.
NCLDV
41
ssDNA bacteriophage;has only a few genes and shows the phenomenon of overlapping genes; contains a circular genome of 5386 nucleotides inside a tiny icosahedral virion, about 25 nm in diameter
Bacteriophage phiX174
42
ssDNA bacteriophage; a filamentous virus with helical symmetry; the virion is long and thin and attaches to the pilus of its host cell; like other filamentous virus it has the unusual property of being released from the host cell without the cell undergoing lysis;
Bacteriophage M13
43
dsDNA bacteriophage; a large, tailed, double-stranded DNA virus of Escherichia coli that possesses complex morphology; kills it host; differs from many other bacteriophage genomes in that its DNA contains the modified cytosine base 5-hydroxymethylcytosine in place of cytosine; encodes its own DNA polymerase; although each copy of the genome contains the same set of genes, they are arranged in a different order (circular permutation);
bacteriophage T4
44
When the T4 DNA is packaged into capsids, the concatemer is not cut at a specific sequence; instead linear segments of DNA just long enough to fill a phage head are generated.
This is called headful packaging and is common among bacteriophages.
45
dsDNA bacteriophage;a relatively small virulent DNA virus that infects Escherichia coli and a few related enteric bacteria. The virion has an icosahedral head and a very short tail; RNA polymerase recognizes only its gene promoters distributed along the its genome; endonuclease cuts the concatemer at specific sequences thus, the DNA sequence in each virion is identical.
Bacteriophage T7
46
dsDNA bacteriophage; also infects Escherichia coli, is a double-stranded DNA virus with a head and tail; a temperate bacteriophage
lambda
47
lambda regulation; represses the transcription of all other lambda-encoded genes, including Cro. When this happens, the lambda genome integrates into the host’s genome and becomes a prophage
accumulation of cI
48
lambda regulation;represses expression of a protein called cII whose function is to activate the synthesis of cI. Hence, following infection, if cI is present at insufficient levels to repress expression of phage specific genes, it accumulates in the cell; if this happens, lambda travels the lytic pathway.
Cro
49
lambda regulation; function is to stabilize cII and protect it from protease attack
cIII
50
DNA Archaeal viruses; virus morphology that commonly affects Euryarchaeota
head and tail type
51
DNA Archaeal viruses; The most morphologically distinctive archaeal viruses infect what phyla
hyperthermophilic Crenarchaeota
52
DNA Archaeal viruses; infects crenarchaeota; forms spindle-shaped virions that often cluster in rosettes; also forms a rigid, helical rod-shaped structure
SSV, SIFV
53
DNA Archaeal viruses; forms a rigid, helical rod-shaped structure; A spindle-shaped virus that infects the hyperthermophile Acidianus displays a novel behavior. The virion contains a circular genome of about 68 kilobase pairs and is lemon-shaped when first released from the host cells. However, shortly after release from its lysed host cell, the virion produces long, thin tails, one at each end
ATV
54
DNA Archaeal viruses;A spindle-shaped virus also infects the hyperthermophile Pyrococcus (Euryarchaeota); resembles SSV but is larger and contains a very short tail and do not have common evolutionary roots;
PAV1
55
ssDNA Archaeal viruses; infects the extreme halophile Halorubrum and is unusual not only because of its circular single-stranded DNA genome but also due to its enveloped nonuniform morphology
Pleomorphic virus 1 (HRPV-1)
56
ssDNA Archaeal viruses; infects the hyperthermophile Aeropyrum pernix . A filamentous capsid surrounds this circular single stranded 24.9-kilobase viral genome, which is double the size of the second largest known single-stranded DNA viral genome.
Aeropyrum coil-shaped virus (ACV)
57
archaea unique method of viral release; their genomes encode structural proteins for their capsids, they also encode a protein called PVAP (protein forming virus-associated pyramid) that facilitates viral release from the cell. PVAP assembles into seven-fold pyramid structures that rupture the host cell’s S-layer and open outwards to create a portal for viruses to exit the host cell.
S. islandicus rod-shaped virus 2 (SIRV2) and turreted icosahedral virus (STIV)
58
uniquely replicating DNA animal virus; first virus to be studied in any detail and was the first virus for which a vaccine was developed; among the largest of all viruses, the brick-shaped vaccinia virions measuring almost 400 nm in diameter; replication occurs in cytoplasm
pox virus
59
uniquely replicating DNA animal virus; group of small and naked icosahedral viruses that contain linear double-stranded DNA genomes; Attached to the 5′ end of the genomic DNA is a protein called the adenoviral terminal protein, and it is essential for replication of the genome. The complementary DNA strands also have inverted terminal repeats that play a role in the replication process; of minor health importance, causing mild respiratory infections in humans; dsDNA is replicated without the formation of a lagging strand
adenoviruses
60
DNA tumor viruses; naked icosahedral virus that can cause tumors in small mammals, such as hamsters and rats. Its circular genome consists of double-stranded DNA; genome is too small (5.2 kb) to encode its own DNA polymerase, so host DNA polymerases are used and DNA is replicated in a bidirectional fashion from a single origin of replication; When SV40 infects a host cell, one of two outcomes can occur, In permissive hosts, virus infection results in the usual formation of new virions and the lysis of the host cell. In nonpermissive hosts, lytic events do not occur; instead, the viral DNA becomes integrated into host DNA, genetically altering the cells in the process; allowing cells to be malignant; transcription of the viral genome occurs in nucleus
Polyomavirus SV40
61
DNA tumor viruses; a large group of double-stranded DNA viruses that cause a variety of human diseases, including fever blisters (cold sores), venereal herpes, chicken pox, shingles, and infectious mononucleosis; examples- Epstein–Barr virus causes Burkitt’s lymphoma, CMV; remain latent in the body for long periods and become active under conditions of stress or when the immune system is compromised; enveloped; replication in the nucleus
Herpesviruses
62
pos strand RNA viruses; The virus infects cells of Escherichia coli by attaching to the cell’s pilus; encodes only four proteins, including the maturation protein, coat protein, lysis protein, and one subunit of RNA replicase, the enzyme that replicates the viral RNA
Bacteriophage MS2
63
pos strand RNA viruses; one of the smallest of all viruses; genome (about 7.4 kb) is also the mRNA, and the VPg protein facilitates binding of the RNA to host ribosomes; replication in host cell cytoplasm
poliovirus
64
pos strand RNA viruses; single-stranded plus RNA viruses that; replicate in the cytoplasm; cause respiratory infections in humans and other animals, including about 15% of common colds; enveloped and contain club-shaped glycoprotein spikes on their surfaces; the largest of any known RNA viruses, about 30 kb; virions are assembled within the Golgi complex,
coronaviruses
65
neg strand RNA Animal Virus; causes the fatal neuroinflammatory disease rabies ;is a rhabdovirus (rod-shaped); genome transcribed by replicase in the cytoplasm; with nucleocapsid and envelope
rabies virus
66
neg strand RNA Animal Virus; an enveloped virus in which the viral genome is present in the virion in a number of separate pieces, a condition called a segmented genome (8 linear single stranded molecules; pleiomorphic due to budding; with hemagglutinin and neuraminidase; also rna replicase and rna endonuclease; exhibits antigenic shift in which segments of the RNA genome from two different strains of the virus infecting the same cell are reassorted generating hybrids that express unique surface proteins unrecognized by the immune system
influenza virus
67
dsRNA virus; a typical reovirus and is the most common cause of diarrhea in infants 6 to 24 months of age; segmented genome
rotavirus
68
dsRNA virus; replication occurs exclusively in the host cytoplasm but within the nucleocapsid itself; RNA replication is conservative as only the minus strand is a template in the infecting nucleocapsids, whereas only the plus strand is a template in the synthesis of double-stranded genomic RNA from assimilated plus-strand RNA copies in progeny virions
reovirus
69
viruses that uses reverse transcriptase; have enveloped virions that contain two identical copies of the single-stranded plus complementarity RNA genome; name refers to the fact that these viruses transfer information from RNA to DNA; To begin the process, the genome is converted to DNA by reverse transcriptase inside the nucleocapsid and then the DNA is released to the cytoplasm
retrovirus; HIV
70
viruses that uses reverse transcriptase; tiny DNA genomes re unusual because they are neither single-stranded nor double-stranded; instead, they are partially double-stranded.
hepadnaviruses; HBV
71
subviral agents; are infectious RNA molecules that lack a protein component; small, circular, single-stranded RNA molecules that are the smallest known pathogens; cause a number of important plant diseases and can have a severe agricultural impact(ie coconut cadang-cadang viroid); infects plants; known to yield small interfering RNAs (siRNAs) as a by-product of replication
viroids
72
subviral agents; infectious agents whose extracellular form consists entirely of protein; lacks both DNA and RNA; cause several neurological diseases such as scrapie in sheep, bovine spongiform encephalopathy (BSE or “mad cow disease”) in cattle, chronic wasting disease in deer and elk. and kuru and variant Creutzfeldt–Jakob disease in humans by catalyzing protein conformational changes that lead to protein clumping and accumulation; none in plants some in yeasts; forms amyloids;
prions
73
subviral agents; nonpathogenic prions; a protein that regulates the transcription of genes encoding certain nitrogen metabolism functions
[URE3] prion
74
subviral agents; nonpathogenic prions; s a human protein that is part of our innate immune system and has been shown to convert to a self-perpetuating prion-like form in cells that become infected with a virus. its aggregation triggers the production of immune modulators called interferons;
MAVS protein
75
valuable tools to visualize the localization of specific proteins and to monitor gene expression in cells
reporter genes
76
encode proteins that are easy to detect or assay and are used by fusing them to genes of interest in such a way that both the reporter gene and the gene of interest are coexpressed
reporter genes
77
For visualizing molecular events, reporter genes that encode fluorescent products such as the
green fluorescent protein (GFP)
78
leads to unwinding of the DNA and loading of the replisome for chromosome replication in Escherichia coli
binding of DnaA to specific DNA sequences within the oriC region of the chromosome
79
one of several penicillin-binding proteins present in the cell.
FtsI
80
ensures the divisome forms only at the center and not at the cell poles
Min proteins (MinC, MinD, MinE)
81
controls the assembly site of FtsZ in Caulobacter
MipZ ( midcell positioning of FtsZ)
82
The major shape-determining factor in Bacteria is the protein
MreB
83
Inactivation of the gene encoding MreB or other proteins in the elongasome leads
rod-shaped bacteria to become coccus-shaped.
84
In the vibrio-shaped bacterium Caulobacter, a shape-determining protein is present in addition to MreB called
crescentin
85
a plant pathogen, elongates at one pole in a process called unipolar elongation
Agrobacterium tumefaciens
86
insertion of new peptdidoglycan; plays a major role in the simultaneous insertion of peptidoglycan precursors.
bactoprenol
87
insertion of new peptdidoglycan; a hydrophobic C55 alcohol that is bonded to an N-acetylglucosamine/N-acetylmuramic acid/ pentapeptide peptidoglycan precursor to form lipid II
bactoprenol
88
insertion of new peptdidoglycan; ??? transports peptidoglycan precursors across the cytoplasmic membrane by rendering lipid II sufficiently hydrophobic to pass through a transmembrane ABC transporter known as ???
bactoprenol, flippase
89
insertion of new peptdidoglycan; lipid II interacts with peptidoglycan polymerases called ??? that insert peptidoglycan precursors into a growing point in the cell wall and catalyze glycosidic bond formation (the remaining bactoprenol is then recycled to the cytoplasm to begin a new round of precursor transport)
transglycosylases
90
insertion of new peptdidoglycan; prior to insertion of the peptidoglycan precursor, small gaps in the existing peptidoglycan are made by enzymes called; enzymes that function to hydrolyze the bond that connects N-acetylglucosamine with N-acetylmuramic acid in the peptidoglycan backbone.
autolysins
91
insertion of new peptdidoglycan; final step in peptidoglycan synthesis; forms the peptide cross-links between muramic acid residues in adjacent glycan chains
transpeptidation
92
Endospore formation sporulation factors; When highly phosphorylated, sporulation proceeds; controls the expression of several sporulation-specific genes
Spo0A
93
Triggers for Activating Endospore Germination; receptors present in the endospore
germinant receptors (GRs)
94
Triggers for Activating Endospore Germination; GRs in a cluster which functions to sense and bind nutrients such as amino acids, sugars, and cell wall peptides from vegetative cells.
germinosome
95
Triggers for Activating Endospore Germination; next step to spore germination after stage I
triggering of cortex lytic enzymes (CLE) to degrade peptidoglycan in the endospore cortex,
96
Triggers for Activating Endospore Germination; modified from normal peptidoglycan such that CLE can recognize it and not degrade cell wall peptidoglycan produced during outgrowth of vegetative cells.
cortex peptidoglycan
97
Triggers for Activating Endospore Germination; major event of stage II- germination
removal of he endospore cortex
98
Triggers for Activating Endospore Germination ; triggered by the increased expression of genes encoding enzymes that biosynthesize peptidoglycan, teichoic acid and lipoteichoic acids, and the cell division protein FtsZ
stage III- outgrowth
99
Caulobacter Differentiation; three major regulatory proteins whose concentrations oscillate in succession.
GcrA, CtrA, DnaA
100
Caulobacter Differentiation; the transcriptional regulators
GcrA and CtrA
101
Caulobacter Differentiation;a protein that functions both in its normal role in initiating DNA replication and also as a transcriptional regulator
DnaA
102
Anabaena heterocyst; The cascade of events leading to heterocyst formation is triggered by
a limitation of “fixed” nitrogen (nitrate, ammonia, etc.)
103
Anabaena heterocyst; limitation of “fixed” nitrogen (nitrate, ammonia, etc.) is sensed in the vegetative cell as an elevation in levels of ???? , the acceptor molecule for formation of the amino acid glutamate
a-ketoglutarate
104
Anabaena heterocyst; the major transcriptional regulator controlling heterocyst formation
HetR
105
Biofilm formation; The actual switch from planktonic to biofilm growth in many bacteria is triggered by the cellular accumulation of the regulatory nucleotide
cyclic di-guanosine monophosphate (c-di-GMP)
106
Biofilm formation; a classic opportunistic pathogen and from its primary reservoir in soil can infect the blood, lungs, urinary tract, ears, skin, and other tissues of humans; causes The major symptoms of the human genetic disease cystic fibrosis; form a tenacious biofilm containing specific polysaccharides that subsequently increase its pathogenicity and prevent the penetration of antibiotics
Pseudomonas aeruginosa
107
Biofilm formation; regulatory molecules
acyl homoserine lactones (AHLs)
108
Biofilm formation; Elevated c-di-GMP levels initiate the production of EPS, including a polysaccharide called which functions as both a primary scaffold for the microbial community and a mechanism for resisting the penetration of antibiotics
Pel
109
Biofilm formation; archaea that forms biofilm
motile hyperthermophilic sulfur chemolithotroph Sulfolobus acidocaldarius
110
Antibiotics; target DNA gyrase in gram-negative bacteria and topoisomerase IV in gram-positive bacteria; lead to cell death by interfering with DNA unwinding and replication
quinolones such as ciprofloxacin
111
Antibiotics; prevent RNA synthesis by either blocking the RNA polymerase active site or blocking RNA elongation by binding to the major groove in DNA
rifampin and actinomycin
112
Antibiotics; contains a region that mimics the 3′ end of a tRNA, and this structural mimicry results in specific binding of the antibiotic to the A site in the 70S ribosome; this induces chain termination and effectively shuts down protein synthesis.
puromycin
113
Antibiotics; specifically target the 16S rRNA of the 30S ribosome and result in the ribosome misreading mRNAs, thus leading to error-filled proteins that accumulate in the cell and ultimately inhibit growth.
Aminoglycoside antibiotics such as streptomycin
114
Antibiotics; a lipopeptide produced by certain Streptomyces species that specifically binds to phosphatidylglycerol residues of the bacterial cytoplasmic membrane; this leads to pore formation and depolarization of the membrane, ultimately resulting in cell death.
Daptomycin
115
Antibiotics; cyclic peptides whose long hydrophobic tails specifically target the LPS layer and ultimately disrupt membrane structure, causing leakage and cell death. (gneg)
polymyxins
116
Antibiotics; target the synthesis of peptidoglycan in bacteria; inhibit growth by interfering with proteins that catalyze transpeptidation
b-lactams penicillin, cephalosporin, and their derivatives
117
Antibiotics; inhibits peptidoglycan synthesis in gram-positive bacteria by binding to the pentapeptide of peptidoglycan precursors and preventing the formation of peptide interbridges by transpeptidases.
vancomycin
118
Antibiotics; prevents peptidoglycan synthesis by binding to the peptidoglycan precursor transport system (bactoprenol) and preventing new peptidoglycan precursors from reaching the site of peptidoglycan synthesis. As autolysins continue to introduce small gaps in the existing peptidoglycan, a shortage of precursors to patch the gaps weakens the cell wall and leads to cell lysis
topical antibiotic bacitracin
119
bacterial resistance mechanisms to antibiotics
(1) modification of the drug target, (2) enzymatic inactivation, (3) removal from the cell via efflux pumps, and (4) metabolic bypasses
120
exhibits antibiotic resistance by producing an alternative penicillin-binding protein called MecA that is unaffected by methicillin or other b-lactam antibiotics.
Methicillin-resistant Staphylococcus aureus
121
mechanisms behind MRSA
presence of the Staphylococcus chromosomal cassette for methicillin resistance (SCCmec) containing the gene for MecA, which is activated in the presence of methicillin or other b-lactam antibiotics antibiotics through a regulatory mechanism involving MecI and MecR1 proteins.
122
eoccurs when a population of antibiotic-sensitive bacteria produces rare cells that transiently become tolerant to multiple antibiotics
persistence
123
genes that encode two components: a toxin whose production inhibits cell growth and an antitoxin that counteracts the activity of the toxin.
Toxin–antitoxin (TA) modules
124
encode a TA module that has been shown to trigger persistence in E. coli
hipAB genes; HipA is a toxin that inhibits translation and HipB is an antitoxin that is susceptible to a protease called Lon.
125
a heritable change in the base sequence of that genome, that is, a change that is passed from the mother cell to progeny cells
mutation
126
a strain of an organism or a virus isolated from nature is called the
wild-type strain
127
A cell or virus derived from the wild type whose genome carries a change in nucleotide sequence is called a
mutant
128
A mutant by definition differs from the wild-type strain in its
genotype
129
nutritionally defective mutants can be detected by the technique of
replica plating
130
A mutant strain with an additional nutritional requirement above that of the wild type or parental strain from which it was derived is called an
auxotroph
131
the strain from which an auxotroph originates is called a
prototroph
132
mutations that occur without external intervention, and most result from occasional errors in the pairing of bases by DNA polymerase during DNA replication
spontaneous mutation
133
mutations that are e caused by environmental agents and include mutations made deliberately by humans.
induced mutations
134
Mutations that change only one base pair and occur when a single base-pair substitution occurs in the DNA
point mutations
135
a change in the RNA from UAC to UAU would have no apparent effect because UAU is also a tyrosine codon. Although they do not affect the sequence of the encoded polypeptide, such changes in the DNA are considered one type of mutation, that is, a mutation that does not affect the phenotype of the cell.
silent mutation
136
a single base change from UAC to AAC results in an amino acid change within the polypeptide from tyrosine to asparagine at a specific site; the informational “sense” (the precise sequence of amino acids) in the polypeptide has changed. this is called a
missense mutation
137
Another possible outcome of a base-pair substitution is the formation of a stop codon in the DNA. This results in prematture termination of translation, leading to an incomplete polypeptide
nonsense mutation
138
are mutations in which one purine base (A or G) is substituted for another purine, or one pyrimidine base (C or T) is substituted for another pyrimidine
transitions
139
point mutations in which a purine base is substituted for a pyrimidine base, or vice versa.
transversions
140
any deletion or insertion of a single base pair results in a shift in the reading frame; often have serious consequences
frameshift mutations
141
many large insertion mutations are due to the insertion of specific DNA sequences called
transposable elements
142
Point mutations are typically reversible, a process known as
reversion
143
a strain in which the original phenotype that was changed by mutation is restored by a second mutation.
revertant
144
the mutation that restores activity is at the same site as the original mutation
same-site revertants
145
If the back mutation is not only at the same site but also restores the original sequence, it is called a
true revertant
146
the mutation is at a different site in the DNA; can restore a wild-type phenotype if they function as suppressor mutations—mutations that compensate for the effect of the original mutation
second-site revertants
147
class of suppressor mutations
(1) a mutation somewhere else in the same gene that restores enzyme function, such as a second frameshift mutation near the first that restores the original reading frame ; (2) a mutation in another gene that restores the function of the original mutated gene; and (3) a mutation in another gene that results in the production of an enzyme that can replace the nonfunctional one.
148
For most microorganisms, errors in DNA replication occur at a frequency of
10-6 to 10-7 per thousand bases during a single round of replication
149
Single base errors during DNA replication are more likely to lead to missense mutations than to nonsense mutations because
most single base substitutions yield codons that encode other amino acids
150
a variety of chemical, physical, and biological agents that can increase the mutation rate and are therefore said to induce mutations.
mutagens
151
molecules that resemble the purine and pyrimidine bases of DNA in structure but display faulty base-pairing properties; DNA may replicate normally most of the time however, DNA replication errors occur at higher frequencies at these sites due to incorrect base pairing. The result is the incorporation of a mismatched base into the new strand of DNA and thus introduction of a mutation; mutation is revealed during the subsequent segregation of this strand in cell division
nucleotide base analogs
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powerful mutagens and generally induce mutations at higher frequency than base analogs; chemical mutagens induce chemical modifications in one base or another, resulting in faulty base pairing or related changes
alkylating agents (chemicals that react with amino, carboxyl, and hydroxyl groups by substituting them with alkyl groups) such as nitrosoguanidine
153
chemical mutagens; are planar molecules that function as intercalating agents; inserted between two DNA base pairs and push them apart. Then, during replication, this abnormal conformation can trigger single base insertions or deletions. Thus, typically induce frameshift rather than point mutations; Ethidium bromide
acridines
154
e two forms of electromagnetic radiation that are highly mutagenic
Nonionizing and ionizing radiation
155
widely used to generate mutations because the purine and pyrimidine bases of nucleic acids absorb UV radiation strongly (the absorption maximum for DNA and RNA is at 260 nm). The primary mutagenic effect is the production of pyrimidine dimers, in which two adjacent pyrimidine bases (cytosine or thymine) on the same strand of DNA become covalently bonded to one another
UV radiation
156
These rays cause water and other substances to ionize, resulting in the formation of free radicals such as the hydroxyl radical that can damage macromolecules in the cell, including DNA. This causes double-stranded and single-stranded breaks that may lead to rearrangements or large deletions.
ionizing radiation x-rays, cosmic rays, gamma rays
157
system initiates a number of DNA repair processes, some of which are error-free; also allows DNA repair to occur without a template, that is, with random incorporation of nucleotide precursors (deoxyribonucleotide triphosphates [dNTPs]); results in many errors and hence many mutations; mutations induced are better than the alternative (death of the cell), as mutations can often be corrected while chromosome breaks usually cannot.
SOS repair system
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In DNA damage tolerance, DNA lesions remain in the DNA, but are bypassed by specialized DNA polymerases that can move past DNA damage—a process
translesion synthesis
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In E. coli, in which the process of mutagenesis has been studied in great detail, the two error-prone repair polymerases are
DNA polymerase V, an enzyme encoded by the umuCD genes, and DNA polymerase IV, encoded by dinB Both are induced as part of the SOS repair system
160
The master regulators of the SOS system are the proteins
LexA and RecA.
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SOS repair system; a repressor that normally prevents expression of the SOS system.
LexA
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SOS repair system; normally functions in genetic recombination , is activated by the presence of DNA damage, in particular by the single-stranded DNA that results when replication stalls.
RecA
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It should be noted that DNA transfer typically occurs in only one direction, from
donor to recipient
164
DNA that enters the cell by horizontal gene transfer faces three possible fates:
(1) It may be degraded by the recipient cell’s restriction enzymes or other DNA destruction systems (2) it may replicate by itself (but only if it possesses its own origin of replication, such as a plasmid or phage genome); or (3) it may recombine with the recipient cell’s chromosome.
165
is the physical exchange of DNA between genetic elements (structures that carry genetic information).
recombination
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This type of recombination is often essential to a cell’s ability to retain DNA following genetic exchange and is behind the well-known phenomenon of “crossing over” in the genetics of eukaryotes.
homologous recombination
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key to homologous recombination.
RecA protein (SOS repair system)
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if a copy of the wild-type gene is introduced into the same cell on a plasmid or viral genome, this gene will encode the necessary protein and, assuming the gene is transcribed and translated, will restore the wild-type phenotype. This process is called
complementation because the wild-type gene is said to complement the mutation, in this case converting the Trp- cell into Trp+
169
DNA derived from virtually any portion of the host genome is packaged inside the mature virion in place of the virus genome.
generalized transduction
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DNA from a specific region of the host chromosome is integrated directly into the virus genome—usually replacing some of the virus genes. This occurs only with certain temperate viruses such as phage lambda
specialized transduction
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during lytic infection, the enzymes responsible for packaging viral DNA into the bacteriophage sometimes package host DNA accidentally. The result is called a
transducing particle
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allows the transfer of any gene from one bacterium to another, but at a low frequency.
Generalized transduction
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allows extremely efficient transfer but is selective and transfers only a small region of the bacterial chromosome
specialized transduction
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Alteration of the phenotype of a host cell by lysogenization is called
phage conversion
175
are the result of prokaryotic cells hijacking defective viruses and using them specifically for DNA exchange; defective bacteriophages
gene transfer agents (GTAs)
176
The F plasmid is actually an episome, a plasmid that
can integrate into the host chromosome.
177
Cells possessing a nonintegrated F plasmid are called
F+
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F plasmid integrated into the chromosome are called
Hfr
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Overall, the presence of the F plasmid results in three distinct changes in a cell:
(1) the ability to synthesize the F pilus (2) the mobilization of DNA for transfer to another cell, and (3) the alteration of surface receptors so the cell can no longer be a recipient in conjugation and is therefore unable to take up a second copy of the F plasmid or any genetically related plasmids.
180
F plasmids containing chromosomal genes are called
F′ (F-prime) plasmids.
181
used to inhibit growth of extreme halophiles,
novobiocin (a DNA gyrase inhibitor) and mevinolin (an inhibitor of isoprenoid biosynthesis)
182
inhibit methanogens
puromycin and neomycin (both protein synthesis inhibitors)
183
mimics a virus because the genetic element encodes a portion of a structurally defined particle that transfers the plasmid between Halorubrum lacusprofundi strains.
pR1SE
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Only one archaeal virus, which infects the thermophilic methanogen ???, has been shown to transduce host genes.
Methanothermobacter thermautotrophicus
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stretches of DNA that can move from one site to another.
transposable elements
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The two major types of transposable elements in Bacteria are
insertion sequences (ISs) and transposons
187
the simplest type of transposable element. They are short DNA segments, about 1000 nucleotides long, and typically contain inverted repeats of 10–50 base pairs.
insertion sequences (IS)
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larger than IS elements but have the same two essential components: inverted repeats at both ends and a gene encoding the transposase
transposons
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transposon which encodes resistance to the antibiotics kanamycin (and neomycin), streptomycin, and bleomycin
transposon Tn5
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transposon which encodes tetracycline resistance.
Tn10
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Two mechanisms of transposition are known:
conservative and replicative
192
as occurs with the transposon Tn5 , the transposon is excised from one location and reinserted at a second location. The copy number of a conservative transposon therefore remains at one
conservative transposition
193
a new copy of the transposon is produced and inserted at the second location. Thus, after this transposition event, one copy of the transposon remains at the original site, while a second copy is incorporated at the new site.
replicative transposition
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A major antiviral defense system present in both Bacteria and Archaea is ??? which functions to seek out and destroy foreign nucleic acid.
CRISPR—clustered regularly interspaced short palindromic repeats
195
These proteins possess endonuclease activity and both mediate the defense against foreign DNA and incorporate new spacers into the CRISPR region.
Cas proteins (CRISPR- associated)
196
correspond to sequences of viral or other foreign DNA and function as a “memory bank” of past encounters with a virus in a manner reminiscent of how animals produce antibodies and long-lasting memory cells against and infecting virus; alternate with CRISPR regions are short repeats of constant DNA sequence present on the host chromosome
spacers
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the first ammonia-oxidizing (nitrifying) archaeon known,
Nitrosopumilus,
198
refers to the use of computers to store and analyze the sequences and structures of nucleic acids and proteins
bioinformatics
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refers to the use of computers to store and analyze the sequences and structures of nucleic acids and proteins
dideoxy method developed by the British scientist Fred Sanger
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second-generation sequencing method still widely used today, is based on the Sanger method and employs the light-emitting enzyme luciferase to detect incorporation of dNTPs by emitting a pulse of light
pyrosequencing
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in Bacteria, translation begins at start codons located immediately downstream of a ribosome-binding sequence (RBS ) on the mRNA also known as
Shine–Dalgarno site
202
some codons are used more frequently than others. The is known as
codon bias
203
is the science that analyzes pooled DNA or RNA from an environmental sample containing organisms that have not been isolated and identified
metagenomics
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Just as the total gene content of an organism is its genome, so the total gene content of a microbial community is its
metagenome
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When DNA is denatured (that is, the two strands are separated), the single strands can form hybrid double-stranded molecules with other nucleic acid molecules by complementary or almost complementary base pairing
hybridization
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The single-stranded segments of nucleic acid, whose identity is already known, are called
nucleic acid probes
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The genome-wide study of the structure, function, and activity of an organism’s proteins is called
proteomics
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is an advanced version of mass spectrometry that does not require the protein separation and digestion step; Instead the sample is affixed to a matrix and then ionized and vaporized by a laser
MALDI (matrix-assisted laser desorption ionization)
209
ons produced in MALDI are accelerated towards the detector by an electric field, where the time of flight (TOF) of each ion is measured. The smaller the mass/charge ratio of an ion, the faster it moves. The detector records the TOF of each ion, allowing the computer to calculate the mass and molecular formula.
MALDI-TOF
210
a more useful technique that can directly analyze biological samples without the need for special analytical preparation;a laser is used to ionize the samplebut the silicon-coated surface used in NIMS does not generate the background interference seen during ionization of a matrix by MALDI-TOF
nanostructure-initiator mass spectrometry (NIMS)
211
critical for studyingthe metabolic potential of microorganisms in natural microbial communities
single-cell genomics (SCG)
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Sequencing DNA from single cells relies on a modified version of the polymerase chain reaction called
multiple displacement amplification (MDA)
213
refers to the use of in vitro techniques to alter genes in the laboratory.
Genetic engineering
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Taq polymerase, a DNA polymerase isolated from the thermophilic hot spring bacterium is stable to 95°C and thus is unaffected by the denaturation step employed in the PCR
Thermus aquaticus
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A DNA polymerase , Pfu polymerase, from a hyperthermophilic species of Archaea with a growth temperature optimum of 100°C; more thermostable than taq; has proofreading activity
Pyrococcus furiosus
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a technique that employs an agarose gel to separate nucleic acid fragments based on differences in their size and charge
gel electropheresis
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f nucleic acid fragments of known sizes
ladder
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Single-stranded nucleic acids whose identity is already known and that are used in hybridization are called
nucleic acid probes
219
When DNA is denatured, the single strands can be used to form hybrid double-stranded molecules with other single-stranded DNA (or RNA) molecules by complementary base pairing in a process called
nucleic acid hybridization
220
probes of known sequence are hybridized to target DNA fragments that have been separated by gel electrophoresis. The hybridization procedure in which DNA is the target sequence in the gel, and RNA or DNA is the probe, is called a
southern blotting
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uses RNA as the target sequence and DNA or RNA as the probe to detect gene expression.
northern blotting
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approach allows the identification of pathogens in clinical samples or bacteria of interest in environmental samples
fluorescence in situ hybridization (FISH) technique
223
The movement of desired genes from their original source to a small and manipulable genetic element (the vector) is called
molecular cloning
224
The cell is protected from its own restriction enzyme(s) by
hemical modification (typically by methylation) of one of the bases in any potential restriction sites that exist in its genome
225
This restriction enzyme makes staggered cuts, leaving short, single-stranded overhangs called “sticky” ends at the termini of the two fragments
EcoRI
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This restriction enzyme cut both strands of the DNA directly opposite each other, resulting in blunt ends
EcoRV
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an enzyme that covalently links the strands of the vector and the source DNA; If the source DNA is PCR generated, it is used to join the amplified DNA to specialized vectors
DNA ligase
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limitation of cloning with restriction enzymes
modifications can only be made at restriction enzyme sites.
229
Through this method, foreign DNA can be inserted into vectors (or the chromosome) by designing the insert DNA to possess short regions (30–50 bases) of sequence homology to the target DNA molecule and activating recombinase enzymes.
recombineering (recombination-mediated genetic engineering)
230
a standard cloning plasmid that contains an ampicillin resistance gene for selection and a blue–white color-screening system to select for recombinants. It also contains a short segment of artificial DNA containing cut sites for many different restriction enzymes. This segment, called a multiple cloning site (MCS), is inserted into the lacZ gene encoding the lactose-degrading enzyme b-galactosidase
plasmid pUC19
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colorless, can be cleaved by b-galactosidase to generate a blue product in screening tests
X-gal (5-bromo-4-chloro-3-indolyl-b-d-galactopyranoside)
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vectors can replicate and be stably maintained in two distinct organisms, such as E. coli and yeast or E. coli and mammalian cells;
shuttle vectors
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are linear vectors that replicate in yeast like normal chromosomes but have sites where very large fragments of DNA can be inserted.
yeast artificial chromosomes (YACs)
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Because it is easy to grow and manipulate, the workhorse for cloning in eukaryotic cells is the yeast
saccharomyces cerevisiae
235
However, if cloned gene expression is desired, the outer membrane of this gram-negative bacterium can hinder protein secretion. This problem can be overcome using the gram-positive bacterium as a cloning host
B. subtilis
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are designed to allow the experimenter to control the expression of cloned genes.
expression vectors
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are designed to allow the experimenter to control the expression of cloned genes.
expression vectors
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introduce mutations at random in the DNA of the intact organism
conventional mutagens
239
uses synthetic DNA plus DNA cloning techniques to introduce mutations into genes at precisely determined sites.
site-directed mutagenesis
240
the gene encoding GFP was originally cloned from the jellyfish
Aequorea victoria
241
a deficiency of somatotropin in the body results in
hereditary dwarfism
242
While recombinant DNA can be transformed into plant cells by electroporation or transfection, the plasmid from the gram-negative bacterium Agrobacterium tumefaciens, a plant pathogen, can be used to transfer DNA directly into the cells of certain plants.
Ti plasmid
243
Genetic engineering can modify a pathogen by deleting genes from its genome that encode virulence factors while retaining those whose products elicit an immune response. This yields a recombinant and infective vaccine that is considered
attenuated because it is less virulent than the original strain
244
one can add genes from a pathogenic virus to the genome of a relatively harmless virus, called a carrier virus. Such vaccines are called ??? and induce immunity to the pathogenic virus.
vector vaccines
245
a vaccine that immunizes against two different diseases at the same time.
polyvalent vaccine
246
vaccines that contain only a specific protein or two from a pathogen, are also produced by recombinant means.
subunit vaccines
247
A set of genes that have all descended from a single ancestral gene present in a shared ancestor are
homologs
248
Genes that are highly similar in sequence often have the same function, and homologous genes that share the same function are orthologs.
orthologs
249
Genes that are homologous but have different functions are
paralogs
250
Horizontal gene transfer is facilitated by this; it is the sum total of all mobile genetic elements in a genome.
mobilome
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genome segment; consisting of genes shared by all individual strains of a species
core genome
252
genome segment; consisting of the core genome plus genes that are not shared by all strains of a species
pan genome
253
contain clusters of genes for specialized functions
chromosomal islands (also called genomic islands)
254
Chromosomal islands that encode virulence factors, molecules that facilitate disease and are found in pathogenic bacteria are called
pathogenicity islands
255
SSU rRNA genes remains a cornerstone of molecular phylogeny in microbiology because they are
highly conserved present in all cellular organisms easily sequenced and analyzed
256
Chemotrophic Metabolism; does not require an external electron acceptor and ATP is generated primarily by substrate-level phosphorylation, with electron balance obtained by reducing metabolic intermediates that are then excreted as fermentation products
fermentation
257
Chemotrophic Metabolism;, requires an external electron acceptor, and ATP is generated by oxidative phosphorylation resulting from electron transport reactions that generate a proton motive force (pmf); the enzyme complex ATP synthase harnesses this force to make ATP
respiration
258
The most important assimilative process in the biosphere
CO2 fixation performed by autotrophic organisms
259
most widespread and globally important pathway for CO2 fixation.
Calvin cycle
260
The key enzyme of the Calvin cycle is the enzyme
RuBisCO
261
proteinaceous microcompartments and are the site of RuBisCO activity.
carboxysomes
262
a pathway of CO2 fixation used by green sulfur bacteria such as Chlorobium, many anaerobic and microaerophilic chemolithotrophic Bacteria
The reverse citric acid cycle (also called the reductive tricarboxylic acid cycle or rTCA cycle)
263
rTCA unique ezymes
a-ketoglutarate synthase and pyruvate synthase, which catalyze the reductive fixation of CO2 using electrons supplied by Fdred.
264
TCA enzymes substituted in rTCA
citrate synthase with the enzyme citrate lyase (an ATP-dependent enzyme that cleaves citrate into acetyl-CoA and oxaloacetate) succinate dehydrogenase from the citric acid cycle by the FADH-linked fumarate reductase (an enzyme that forms succinate from fumarate) in the reverse cycle.
265
the most efficient of all CO2 fixation pathways
reductive acetyl-CoA pathway
266
describes phototrophic organisms that consume H2O and produce O2 as a waste product; sometimes H2S to H2
oxygenic photosynthesis
267
describes photosynthetic organisms that do not produce O2
anoxygenic photosynthesis
268
contain magnesium instead of iron at the center of the ring; contain specific substituents on the tetrapyrrole ring and a hydrophobic alcohol that helps anchor it into photosynthetic membranes.
chlorophylls
269
Anoxygenic phototrophs produce this chlorophyll equivalent
bacteriochlorophyll
270
In eukaryotic phototrophs, photosynthesis takes place in intracellular organelles called
chloroplasts
271
chloroplasts, which contain sheetlike photosynthetic membrane systems called
thylakoids
272
The ultimate structure for capturing energy from low light intensities is the
chlorosome
273
The most widespread accessory pigments in phototrophs; hydrophobic pigments that are firmly embedded in the photosynthetic membrane
carotenoids
274
main light-harvesting systems of these phototrophs
phycobiliproteins
275
results in noncyclic photophosphorylation because electrons do not cycle back to reduce the oxidized P680, but instead are used in the reduction of NADP+
oxygenic photosynthesis
276
The reduced inorganic nitrogen compounds ammonia (NH3) and nitrite (NO2 -) are oxidized aerobically by the chemolithotrophic nitrifying bacteria in the process of
nitrification
277
NH3 can also be oxidized under anoxic conditions. This process is called
anammox (for anaerobic ammonia oxidation) and is catalyzed by an unusual group of obligately anaerobic Bacteria.
278
C1 assimilation pathway; acetyl-CoA is synthesized from one molecule of CH2O and one molecule of CO2;
serine pathway
279
c1 assimilation pathway; more energy efficient than the serine pathway because all of the carbon for cell material is derived from CH2O
ribulose monophosphate pathway
280
yields a single fermentation product, lactic acid.
homofermentative
281
yields products in addition to lactate, mainly ethanol plus CO2.
heterfermentative
282
variant of glycolytic pathway; glucose 6-phosphate is oxidized to 6-phosphogluconic acid and NADPH; the 6-phosphogluconic acid is dehydrated and split into pyruvate and glyceraldehyde 3-phosphate (G-3-P)
Entner–Doudoroff pathway
283
three different acids—acetic, lactic, and succinic—are formed from the fermentation of glucose or other sugars that can be converted into glucose. Ethanol, CO2, and H2 are also typically formed as fermentation products;utilize glycolysis
mixed- acid fermentation
284
A number of clostridia ferment sugars, producing ??? and ??? as major fermentation products.
butyric acid, H2