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MICRB 201 > Exam 4 > Flashcards

Exam 4 Flashcards

1
Q

Homologous recombination in Euk.

A

vertical gene transfer during meiosis

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2
Q

Homologous recombination in Prok.

A

horizontal gene transfer by transformation, transduction, conjugation

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3
Q

Genetic recombination

A

the physical exchange of genes between genetic elements

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4
Q

RecA

A

promotes strand invasion

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5
Q

Transformation

A

involves the uptake of naked DNA from environment

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6
Q

Transformation experiment

A

Streptococcus pneumoniae

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7
Q

Streptococcus pneumoniae smooth

A

Capsule and Pathogenic

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8
Q

Streptococcus pneumoniae rough

A

No Capsule and Nonpathogenic

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9
Q

Mechanism of transformation

A

DNA binding proteins expressed on the cell’s surface; stand cleaved by endonucleases (5-15 kilobases); while being taken up, one strand is completely degraded by exonucleases

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10
Q

Competency

A

the ability to take up exogenous DNA; Bacillus, Streptococcus, Haemophilus, Neisseria; Gram +, -, Archaea

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11
Q

Poorly transforming bacteria

A

Electrical shock; exposure to Calcium chloride

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12
Q

Electroporation

A

usually used with Gram +, and then chill them after

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13
Q

Streptococcus pneumoniae competency

A

can take up any DNA; cells must be in exp. Phase of growth; 10^7-8 bact/mL; competence factor secreted; triggers upregulation of genes involved in translation

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14
Q

Bacillus competency

A

only 20% competent after competence factors secreted

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15
Q

Haemophilus competency

A

will only take up its own DNA; an 11 base sequence repeated 1400 times must be recognized

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16
Q

Transduction

A

DNA transfer that is mediated by bacteriophage

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17
Q

lytic

A

generalized transduction

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18
Q

lysogenic

A

specialized transduction

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19
Q

Conjugative plasmids

A

relies on formation of a pili; mediates direct exchange between bacterial cells

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20
Q

tra region

A

a plasmid must have this to transfer DNA

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21
Q

R (resistance) plasmids

A

confer resistance to antibiotics and other growth inhibitors by having genes that encode for proteins that inactivate it

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22
Q

Virulence plasmids

A

carry genes that allow attachment to specific cells and/or the formation of substances that cause damge to host

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23
Q

Conjugation

A

the transfer of DNA by direct cell contact

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24
Q

Pilus

A

thin, fragile, and hard to maintain; retracts for more stability, but still hard to keep in place

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25
Q

Biofilm

A

on the rocks in a stream; a large community of cells create an EXOPOLYMER SUBSTANCE; aids in conjugation

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26
Q

Hfr conjugation

A

chromosomal genes take longer to transfer than plasmids

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27
Q

Transposition

A

recombination due to themovement of DNA sequences form one part of the genome to another; can occur in Euk.; diversifies genome

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28
Q

transposons

A

jumping genes

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29
Q

transposable elements

A

insertion sequences: contain only the genes for those enzymes required for its movement; transposons: carry other genes

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30
Q

Discovery of Viruses

A

A. Mayer was trying to find the cause of Tobacco mosaic: hypothesis- either the organism was ultra-small or toxin mediated

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31
Q

Viruses

A

average size: 10-400nm

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32
Q

Enveloped virus

A

nucleocapsid; main component from host

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33
Q

nucleocapsid

A

main component from host; any proteins within it will be viral

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34
Q

Naked virus

A

lacks a envelope

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35
Q

Glycoprotein spikes

A

used for attachment

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36
Q

capsomeres

A

usually one expressed; MAX 4

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37
Q

Viral capsid structure

A

copmosed of protein subunits called capsomers that spontaneously self assemble

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38
Q

2 Arrangements of capsid structure

A

Helical and Icosahedron

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39
Q

Helical structure

A

rigid or flexible; hollow inside

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40
Q

Icosahedron

A

aka polyhedral capsule; much more common; 20 triangular faces and 12 vertices

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41
Q

Vertices

A

glycoprotein spikes protrude from these in naked viruses

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42
Q

Atypical/complex viruses

A

Pox virus: coating made up of glyco and fibral protein; Bacteriophage

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43
Q

Viral nucleic acid

A

quite small; usually 4-100 genes; can contain either DNA or RNA, NOT BOTH

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44
Q

Viral DNA

A

can be double stranded or single stranded; double more common

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45
Q

Viral RNA

A

can be double stranded or single stranded; single more common

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46
Q

Positive sense RNA

A

can act as mRNA

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47
Q

Negative sense RNA

A

cannot act as mRNA

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48
Q

Segmented viral genome

A

multiple pieces of genome; one gene per piece

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49
Q

Viral Order

A

-virales

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50
Q

Viral Family

A

-viridae

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51
Q

Viral Genus

A

-virus

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52
Q

Viral Species

A

_virus

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53
Q

Viral Taxonomy

A

based on the International Committee for the Taxonomy of Viruses

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54
Q

International Committee for the Taxonomy of Viruses

A

genome structure; life cycle; morphology; genetic relatedness

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55
Q

The Baltimore classifcation system of viruses

A

genome and replication strategy only; 7 classes

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56
Q

Adsorption

A

a specific glycoprotein spike binds to a specific host cell receptor protein

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57
Q

Host cell receptors

A

tend to deal with the immune system; typically cytokine receptors

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58
Q

lipid rafts

A

lipid microdomains that allow only certain proteins to interact; usually have more cholesterol; GPI-anchored protein found here

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59
Q

Penetration and Uncoating: Endocytosis

A

NAKED or ENVELOPED; endocytic entrance; endosomal compartment and a decreease in pH triggers uncoating event

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60
Q

Penetration and Uncoating: Fusion

A

ENVELOPED ONLY; viral envelope directly fused with host cell membrane; uncoating events hard to define

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61
Q

Synthesis: DNA Viruses

A

enter host’s nucleus and exert control over host’s machinery

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62
Q

Why can’t RNA viruses use Euk’s machinery?

A

there is no RNA-dependent RNA polymerase to read the RNA

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63
Q

Synthesis: (-) RNA

A

a viral RNA-dependent RNA polymerase is packaged within the viral capsid

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64
Q

Synthesis: (+) RNA

A

translation of RNA-dependent RNA polymerase is translated by host’s ribosomes and enzymes

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65
Q

Retroviruses

A

RNA is transcribed to DNA using reverse transcriptase; the DS DNA enters the nucleus where it is incorporated into host’s genome using integrase

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66
Q

reverse transcriptase

A

RNA=> DNA

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67
Q

integrase

A

integrates viral DNA into host’s genome

68
Q

Early viral proteins

A

for genome replicationand taking over host cell’s machinery

69
Q

Late viral proteins

A

for capsid

70
Q

matrix proteins

A

line up underneath lipid domains and grab onto nucleocapsid proteins

71
Q

budding

A

reverse endocytosis

72
Q

Cytopathic effects of viruses

A

inhibition of trascription, translation, and host’s replication; endosomes damaged; plasma membrane alterations (syncytia); inclusion bodies formed; build up of of viral products; chromosomal incorporation; malignant cells

73
Q

Syncytia

A

large multi-nucleated cells where the plasma membrane fused together (50-100 nuclei); syncytium (pl)

74
Q

Oncogenes

A

cancer causing genes

75
Q

Proto-oncogenes

A

regulate cell growth and division cycle

76
Q

dsDNA viruses

A

making a viral protein to inhibit a tumor suppresssor gene (pRb, p53)

77
Q

Retroviruses and cancer

A

either bring in their own oncogene protein or insert oncogene into genome

78
Q

Bacteriophage found…?

A

in both bact. and arch.; RNA, ss DNA (not common), dsDNA (common and naked)

79
Q

virulent/lytic

A

kill host immediately (25 minutes)

80
Q

temperent/ lysogenic

A

“hide out” in bacteria

81
Q

Step 1 of Lytic Attachment

A

tail fibers mediate interaction

82
Q

Step 2 of Lytic Attachment

A

retraction of tail fibers pulls base plate clos; lysozyme-like enzyme on base plate breaks peptidoglycan

83
Q

Step 3 of Lytic Attachment

A

tail enters cell wall and contracts

84
Q

Step 4 of Lytic Attachment

A

DNA is released into cytoplasm

85
Q

Early phage PROTEINS

A

involved in DNA replication and transcription

86
Q

Middle phage PROTEINS

A

involved in DNA replication and transcription

87
Q

Late phage PROTEINS

A

involved in the making of head and tail fibers and anything else needed for release

88
Q

Early phage GENES

A

transcribed using host’s RNA polymerase; promoters identical to host

89
Q

Last Early Proteins

A

include an anti-sigma factor; make proteins that modify alpha-subunit to make host RNA polymerase bind to phage promoters; some phage proteins will bind directly to RNA polymerase

90
Q

Anti-sigma factor

A

interacts with sigma-factor and disables it

91
Q

Late phage GENES

A

make a phage sigma-factor that recognizes late gene promoters

92
Q

Phage DNA

A

linear not circular; must overcome th eabsence of telomerase

93
Q

concatomer

A

individual genes spliced together; leaving extra ensures that all genes included

94
Q

Phage endonuclease

A

cuts a “headful” of DNA

95
Q

Phage assembly

A

all parts come from different biosynthetic events and then are assembled

96
Q

Phage release

A

two enzymes are used to release; VERY RARE to see budding or envelopment

97
Q

Holin

A

puts holes in cell membrane for release

98
Q

T4 lysozyme

A

degrades peptidoglycan

99
Q

Characteristics of lysogenic phage

A

can’t have late gene synthesis AND phage genome integrates into host genome

100
Q

Virulent phage

A

can only be lytic

101
Q

Temperent phage

A

can either be lytic or lysogenic

102
Q

Cohesive ends

A

linear genome, but the 5’ ends of each strand has a 12 nucleotide sequence that are complementary

103
Q

cos sites

A

where the single stranded tails bind to make circular genome

104
Q

MOI

A

multiplicity of infection; how many other phage are in the area

105
Q

Host growing rapidly under nutrient rich conditions and has a low MOI

A

Lytic favored

106
Q

Host growing slowly under metabolic stress and high MOI

A

Lysogenic favored

107
Q

If cI wins

A

lysogenic cycle; aka lambda repressor

108
Q

If Cro wins

A

lytic cycle

109
Q

PL

A

makes N protein

110
Q

PR

A

makes Cro

111
Q

N Protein

A

an anti-terminator which allows RNA polymerase to go past specific termination sites to make cII and cIII

112
Q

cII

A

considered unstable, so targeted by bacterial protein hflA; activates transcription of cI

113
Q

hflA

A

degrades cII; available based on how well the cell is doing; cyclic AMP involved

114
Q

cIII

A

stabilizing factor for cII; 2 mechanisms: binds directly to cII and the other is unknown

115
Q

hflA activity when cAMP levels are low in glucose rich environments

A

hflA activity is high

116
Q

hflA activity when cAMP levels are high in glucose lacking environments

A

hflA activity is low

117
Q

Integrase enzyme

A

cleaves phage and host genome at specific sites

118
Q

Can prophage remain integrated indefinitely?

A

Yes

119
Q

Agents that revert phage back to a lytic cycle

A

UV radiation, X-rays, chemicals

120
Q

SOS Response for lytic cycle

A

activates a protease that destroys lambda repressor; lytic genes can now be transcribed

121
Q

Cultivating bacteriophage

A

broth culture containing young, actively growing cells

122
Q

Cultivating animal viruses in whole animals

A

hard to observe effects at cellular level; many viruses only infect one host

123
Q

Cultivating animal viruses in cell culture

A

animal cells can be free from surrounding tissue and grown on plastic to produce a monolayer

124
Q

Primary Cell Lines

A

Straight from the tissue with no passages

125
Q

Continuous Cell Lines

A

Have at least one passage

126
Q

Quantifying viruses

A

differential centrifugation or gradient centrifugation; mixed with liquid agar and a culture of host cells

127
Q

Why are Plaque Forming Units used?

A

cannot be sure if it was one virus or phage that started the colony of death

128
Q

Who discovered Prions?

A

Stanley Prusiner

129
Q

Prion definition

A

Proteinaceous infectious particles found composed of normal protein PrPC

130
Q

Normal PrPC

A

more alpha-helices

131
Q

PrPSc

A

more B-sheets

132
Q

If PrPSc is transmitted by animals

A

infectious ex) Kuru (cannibalism)

133
Q

If PrPSc is sporadic

A

spontaneous random misfolding of PrPC ex) Creutzfeldt-Jacob disease

134
Q

If PrPSc is inherited

A

mutation within the PrP gene (20 mutations linked) ex) Gerstmann-Straussler-Scheinker Disease and Fatal familial insomnia

135
Q

What do prions cause?

A

Transmissable spongiform encephalopathies

136
Q

Amyloids

A

proteinaceous fibers that are resistant to proteases and are an elongation of PrPSc

137
Q

Highest resistance of microbes

A

bacterial endospores

138
Q

Moderate resistance of microbes

A

naked viruses and bacterial cells with more resistant vegetative cells

139
Q

Least resistance of microbes

A

most bacterial vegetative cells, enveloped viruses

140
Q

Moist Heat

A

operates at lower temperates and short exposures; causes coagulation and denaturing of proteins; permeates better using moisture

141
Q

Dry Heat

A

operates at higher temperatures with long exposures; dehydrates cells

142
Q

Autoclave

A

121 degrees Celsius achieved by pressurizing steam by 15psi

143
Q

Traditional pasteurization

A

63 degrees C for 30 mins. BATCH

144
Q

Flash pasteurization

A

71 degrees C for 15 sec. THIN AMOUNT

145
Q

Ultrapasteurization

A

140 degrees C for 1-2 sec. THIN AMOUNT; DOES NOT NEED REFRIGERATION

146
Q

Ionizing radiation

A

causes DNA breakages; Gamm rays, X-rays, Electron beams

147
Q

Nonionizing radiation

A

causes Thymine dimers; Germicidal lamps at 254nm wavelength UV disinfection

148
Q

Advantages of ionization

A

prevent food born illnesses; preservation; control of insects; delay sprouting and ripening; sterilizes food

149
Q

How many KiloGrays of ionization required depending on DNA?

A

the more DNA it has, the less KiloGrays necessary

150
Q

Depth filter

A

liquid and air; 0.3 micrometers = 99.97% of contaminants

151
Q

Membrane filter

A

porous; to be sterilizing 0.2 micrometers

152
Q

Disinfectant

A

for a nonliving surface

153
Q

Antiseptic

A

for a living surface

154
Q

Microbicidal

A

kills organism

155
Q

Microbiostatic

A

inhibits growth

156
Q

What are antimicrobials geared to target?

A

cell wall, cell membrane, genetic material, proteins

157
Q

Which chemical agents come closest to fulfilling all the requirements?

A

Hydrogen Peroxide and Glutaraldehyde

158
Q

Chlorine as a disinfectant

A

hypochlorous acid

159
Q

Iodine as a disinfectant

A

first used as a tincture of 2% iodine and water, ethanol, mix of Potassium idodide; STAINS, ALLERGENIC

160
Q

Iodophores

A

common in use; water soluble; release iodine slowly

161
Q

Phenol

A

Joseph Lister; not widely used because of irritating effects

162
Q

Isopropyl

A

used at 70% isopropyl and 30% water for better permeability

163
Q

Hydrogen Peroxide

A

3% Hyd. Perox. Used; can be vaporized to sterilize enclosed areas

164
Q

Detergents

A

break down cell membrane by interacting with lipids

165
Q

Silver

A

effective in low concentrations

166
Q

Glutaraldehyde

A

used more than formaldehyde; yellow acidic liquid with a mild odor

167
Q

Why do dsDNA phage have to form a concatemer?

A

To overcome the absence of telomerase

MICRB 201 (3 decks)

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