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MOLBIOF!NA1S > trans > Flashcards

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1
Q

_____ – in contrast to bacterial cells, all eukaryotic cells
contain membrane-bound organelles.

A

Organelles

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2
Q

Cell membrane functions as shell of each cell in the body; contains a hydrophilic and hydrophobic
surface; encloses contents of the cell.

A

lipid bilayer

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3
Q

Membrane bound organelles within the cell are encased in a ____

A

lipid bilayer

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4
Q

Structure containing the majority of genetic

material within the cell.

A

nucleus

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5
Q

_____ – genetic material of the cell; organized into

chromosomes.

A

DNA

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6
Q

Ø Numerous within each cell; number varies by cell type.
Ø Energy generation occurs within _____
Ø Oxidative phosphorylation
Ø Small amount of DNA

A

mitochondria

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7
Q

Ø Structure continuous with the nucleus; contains no DNA &

Ø Site of protein synthesis

A

Endoplasmic reticulum

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8
Q

Ø Protein modification &

Ø Protein transport to the cytosol

A

Golgi apparatus

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9
Q

Ø Protein modification &

Ø Protein transport to the cytosol

A

Golgi apparatus

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10
Q

Ø Function in removal of waste products within the cell

A

Lysosomes and peroxisomes

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11
Q

Structure of DNA

a. Purine and pyrimidine bases: A, T, G, C
b. A ______ backbone links the bases together to form a chain.
c. A ______r is also contained at each base position.
d. Double helix. Two DNA chains are _______bonded to one another between A and T or C and G. The two joined, antiparallel strands then twist into a double helix.
e. ______ Along the DNA strands are specific, short regions that encode for ___.

A

phosphodiester
deoxyribose suga
hydrogen
Genes.

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12
Q

Chromosomes – human DNA is further arranged into ___

pairs of matched chromosomes

A

23

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13
Q

– the DNA strands are wound around histone proteins and these coil together to form a tightly packed _____

A

Chromatids

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14
Q

two sister chromatids are joined at

the _____. These two joined chromatids form a chromosome.

A

centromere

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15
Q

the ends of chromosomes contain
repetitive sequences of a specific 6-base sequence: ______. This repeated sequence can be many
hundreds of base pairs long.

_______protect
genes at the ends of chromosomes from normal damage and shortening during DNA replication. They are also thought to be involved in the aging process.

A

TTAGGG

Telomeres

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16
Q

Ø Extrachromosomal, circular pieces of DNA first identified in bacterial cells
Ø Confer antibiotic resistance to bacteria
Ø Engineered in the laboratory to clone specific pieces of human DNA

A

Plasmids

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17
Q

Ø Semiconservative process
Ø Strands are separated by enzymes and new nucleotides added to form a new strand using the original strand as a _____. Replication is carried
out by _______.
Ø Result is the generation of two identical strands of
DNA, each of which contain one strand from the original duplex.
Ø Carried out in the ____ direction. One stand is
directly copied from its template. Other strand is
replicated in small pieces known as Okazaki
fragments. These fragments are later joined by DNA
ligase to form a contiguous strand of DNA.

A

DNA Replication
template
DNA polymerases
3’ to 5’ direction

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18
Q

RNA

  1. Classes
    a. ______ – defines amino acid to be incorporated into a polypeptide
    b. _______ – forms a portion of the ribosome; participates in protein synthesis
    c. ______ – template from which a polypeptide or protein is made
A

tRNA
rRNA
mRNA

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19
Q

_______ – process of converting genetic code contained within the DNA into an RN copy. After this process, the mRNA copy undergoes specific ______ modifications to prevent degradation, function in controlling the rate of protein synthesis, repair misincorporated nucleotides, or increase genetic diversity.

A

Transcription

post-translational

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20
Q

_______ – addition of long sequence of A nucleotides to the 3’ end of mRNA

A

Polyadenylation

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21
Q

addition of a methyl group to the 5’ end of mRNA

A

Capping

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22
Q

______ – inserting, deleting, or editing nucleotides after synthesis

A

splicing

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23
Q

Ø Represented by a triplet codon sequence
Ø Basic components of proteins
Ø Contain an amino and carboxyl group
Ø Twenty _____ found in proteins

A

Amino acids

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24
Q

Ø Chain of amino acids
Ø Alpha-helix or beta-sheet
Ø Component or subunit of a mature protein

A

Polypeptides

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25
Q

Ø All are proteins
Ø Catalyze specific chemical reactions within or outside of the cell
Ø ______ proteins are common examples of enzymes
Ø Degradation of cellular components
Ø Cell division
Ø Also produced commercially for use in the laboratory

A

Enzymes

DNA synthesis proteins

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26
Q

________– most common type of molecular diagnostic specimen. Blood is most commonly collected in ______ tubes. ______ is a potent
inhibitor of PCR and is not used in molecular diagnostics. Blood should be stored at ____ until processing.

A

Whole blood
purple-top
Heparin
4*C

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27
Q

______ – specimens should be frozen immediately upon receipt or held in cell culture medium at _____
until processing

A

Tissue

37*C

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28
Q

______ – often used for remote site testing; collection of DNA specimens for parentage testing.
Swabs should be ____ before shipment to the laboratory.

A

Buccal swabs

air-dried

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29
Q

_____ – urine, feces, sputum, CSF, genital swabs, semen, sections of formalin-fixed paraffinembedded tissues, bone, hair, fingernails usually
confined to ______specimens

A

Other specimens

forensic

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30
Q

DNA isolation

_______ – provide ease of use and high degree of reproducibility. Yields _____amounts of DNA. DNA is usually very pure. Most often used to isolate DNA from blood. This is the most commonly encountered clinical DNA isolation method.

A

Manufacturer (Qiagen) kits

smaller

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31
Q

DNA Isolation

_______– fastest method. Low quality, low mass DNA recovery. Usually used to isolate DNA from buccal/FTA cards.

A

Quick-extract solutions

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32
Q

RNA isolation

a. _______ – gold standard. Yields a large mass of high quality RNA. Can be used
on any specimen type.
b. ________ – provide ease of use and high degree of reproducibility. Preferred method for clinical RNA isolation.
c. ________. Increase recovery of mRNA in specimens.

A

Acid phenol/LiCl salt isolations
Manufacturer kits
Poly-A enrichment kits

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33
Q

Spectrophotometry

Ø ___ value is used to calculate nucleic acid mass.

Ø DNA A260/280 ratio between _______ – indicative of DNA quality sufficient for clinical manipulation. Lower ratios indicate impure specimens.

Ø RNA A260/280 ratio between _____– indicative of RNA quality sufficient for clinical manipulation. Lower ratios indicate ____ specimens.

A

A260
1.8 and 2.0
1.9 and 2.1
impure

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34
Q
  1. Target amplification
    a. _______ – method by which almost all molecular diagnostic assays are performed. It is a three-step process consisting of melting the duplex DNA strands, annealing the primers, and extending the new fragment to yield a specific, short DNA product. This process is repeated 25 to 40 times to theoretically yield trillions of copies
    from one starting copy of DNA.

Ø _____ – short segments of synthetically manufactured DNA used to define a specific
position to be copied in the PCR process. Each PCR reaction requires two primers – one on either side of the fragment to be amplified.

Ø _______ – salt required as a cofactor for the polymerase enzyme used to carry out PCR.

A

Polymerase chain reaction (PCR)
Primers
Magnesium chloride

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35
Q

_______– enzyme used to generate new DNA copies in the PCR process. A thermostable enzyme, meaning that it will not degrade at very high temperatures used in the PCR process.

Ø_____ – containing potassium chloride, sodium chloride, and other additives are also included in the reaction to stabilize the enzymes and DNA during the PCR process.

A

Taq polymerase

Buffers

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36
Q

_______ – individual building blocks of the DNA strand.

A

dNTPs (A, T, G, C)

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37
Q

_______ – a very small mass of this specimen from the patient to be tested is also added to the reaction. The PCR process only requires ______ quantities of input DNA.

A

Genomic DNA

femtomolar

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38
Q

________ – an alternative method to PCR that is not based upon direct amplification of the genomic DNA template.

A

Transcription-based amplification (TMA)

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39
Q

_______ – another method that does not rely on direct amplification of the template DNA strand. Two commonly encountered clinical examples of this strategy
include:

a. ___________
b. _________ – used clinically to detect deletions of multiple entire exons in very long stretches of DNA representing large genes.
c. _____________

A

Probe amplification

Ligase chain reaction
Multiplex ligation-dependent probe amplification (MLPA)
Strand displacement amplification

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40
Q

_______ – another method of detecting a DNA target without direct amplification of the template DNA. It relies on detection of substrates bound to the target nucleic acid.

a. Branched DNA amplification
b. Hybrid capture

A

Signal amplification

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41
Q

Method used in many applications within the molecular diagnostics laboratory. Used to fractionate DNA at sequence-specific locations using restriction enzymes. ______ is used to generate size standards and to prepare genomic DNA for analysis by Southern
blotting and hybridization.

A

Restriction digestion

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42
Q

______– method to analyze DNA contained in an entire genome through a process of restriction digestion, electrophoresis, transfer to a membrane, and hybridization to a sequence-specific probe.

A

Southern blot

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43
Q

_______– analogous to the process of Southern blotting, but is performed on RNA specimens.

A

Northern blot

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44
Q

_______ – small fragments of DNA used to detect a specific gene or mutation in the process of Southern or Northern blotting

A

Probes

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45
Q

_______– conditions, i.e. salt concentration, temperature, time and buffer composition, that control the stringency of the hybridization procedure.

A

Hybridization dynamics

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46
Q

A method used to separate DNA fragments onto a solid matrix based upon size

A

Gel electrophoresis

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47
Q
  1. ___________ – most often performed to resolve DNA fragments larger than 100 bp in size.
  2. __________ – method of choice to resolve small DNA fragments. Very high concentrations of polyacrylamide matrix can resolve down to 1 bp differences in size.
  3. __________ – modification of polyacrylamide gels used in automated DNA sequencing and genotyping platforms.
A

Agarose gel electrophoresis
Polyacrylamide gel electrophoresis
Capillary gel electrophoresis

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48
Q

Quantitative Real-time Polymerase Chain Reaction

A PCR method starting the mRNA as a ____. This method is capable of quantifying the copy number of mRNA transcripts present in a specimen.

  1. ________ – enzyme used to make a copy DNA, or cDNA, from the mRNA transcript contained in a specimen. This process is referred to as cDNA synthesis.
  2. ________– general term for the PCR method used to observe the production of new DNA fragments in real time on an automated platform.

a. _____ – dye often used in quantitative real-time PCR assays to detect newly generated DNA fragments
b. ______ – alternate system of
amplification and detection of fragments in quantitative real-time PCR using dye-labeled probes.

A 
template
Reverse transcriptase
Real-time PCR 
SYBR green
TaqMan chemistry
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49
Q

______ - Allows determination of each base pair in a single strand of DNA.

__________ – original method of DNA sequencing; basis of all DNA sequencing used in the clinical environment today. It is based upon the incorporation of dideoxy nucleotides in random positions in the newly synthesized DNA strand.

_________– method by which DNA
sequence is determined based upon incorporation of a dye-labeled dideoxy nucleotide and resolution on a capillary electrophoresis instrument. Each dideoxy
nucleotide is labeled with a specific dye and when incorporated, is excited by a laser and detected by a CCD
camera on an automated platform.

A

DNA Sequencing

Sanger dideoxy sequencing
Fluorescent DNA sequencing

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50
Q

Technologies are the newest area of use in molecular diagnostics. They fall in two general categories:

  1. ___________ – designed to
    detect single base or other small changes in DNA sequence across the entire genome simultaneously. Many thousands of positions can be interrogated in one experiment.
  2. _________– designed to asses global changes in expression of specific genes in a given tissue at a given time or under a specific set of treatment parameters.
A

Single nucleotide polymorphism arrays

Gene expression arrays

51
Q 
\_\_\_\_\_\_\_\_\_
Ø Trinucleotide repeat expansion
Ø X-linked
Ø Mental retardation
Ø Primary affects males
A

Fragile-X

52
Q

Ø Deletion of entire exons
Ø Dystrophin gene
Ø X-linked
Ø Muscle degeneration

A

Duchenne muscular dystrophy

53
Q

Ø Trinucleotide repeat expansion with anticipation; the worsening of clinical symptoms with increasing repeat size in successive generations
Ø Autosomal dominant
Ø Males and females
Ø Multisystem disease

A

Myotonic dystrophy

54
Q

Ø Many individual mutations in the _____may give rise to CF alone or in combination
Ø ______ mutation accounts for over 70% of CF cases
Ø Autosomal recessive
Although it primarily affects the lungs, it is a multisystem disease
Ø Most common genetic disease in Caucasians

A

CFTR gene

Delta F508

55
Q

DNA isolation

_________ – gold standard. Yields a _______of high quality DNA. Can be used to isolate DNA from any specimen type. This
method normally requires a large mass of starting material.

A

Phenol:Chloroform:Isoamyl alcohol method

large mass

56
Q

_______________ – commonly performed assay in molecular diagnostics. Determination of the effectiveness of a therapy or the tracking of a patient’s disease status based upon the presence of
specific mRNA transcripts known to cause leukemia or
lymphoma.

a. ___________ – presence of
Philadelphia chromosome or 9;22 translocation. The
transcript can be detected by quantitative real-time
PCR and the size determined by fluorescent capillary
electrophoresis.

A

Minimal Residual Disease Detection

Chronic myeloid leukemia (CML )

57
Q

Chronic myeloid leukemia (CML )– presence of
Philadelphia chromosome or 9;22 translocation. The
transcript can be detected by ________ and the size determined by ___________

A

quantitative real-time PCR

fluorescent capillary electrophoresis.

58
Q

_______ – presence of a specific T or B cell clone
originating from a lymphoma. The clone is detected using
_____and _________

A

Clonality

PCR
fluorescent capillary electrophoresis.

59
Q

__________– a genetic phenomenon
observed in cells undergoing changes in specific cells. The
expansion of short repetitive stretches of DNA to much
longer stretches that change gene expression within the
cell.

A

Microsatellite instability

60
Q

______– hereditary form of colon cancer that is

marked by a microsatellite instability

A

HNPCC

61
Q

__________– nature of some tumors to exhibit
loss of function of one allele in a specific gene in a cell in
which the other gene was already inactivated. The most
common example of a cancer caused by this process is
______, in which expression of a tumor
suppressor gene is lost

A

Loss of heterozygosity

retinoblastoma

62
Q

_______________ – can be determined

through genotyping of DNA or gene expression analysis.

A

Drug metabolism and sensitivity

63
Q

____________– performed to determine drug sensitivity. Test is carried out on a microarray platform.

A

Cytochrome P450 genotyping

64
Q

Bacterial identification. The following organisms can be
easily detected,___________identification, using ____:
a. Chlamydia trachomatis
b. Neisseria gonorrheae
c. MRSA
d. VRE

A

qualitative

PCR

65
Q 
Viral identification. Performed in \_\_\_\_\_\_\_\_ manner. 
Following may be \_\_\_\_ detected:
a. Viral load – \_\_\_\_\_
i. HCV
ii. HIV
iii. CV
A

quantitative

66
Q

There are a number of ways by which human identification
may be performed. The most widely used system is based
upon determination of the length of repetitive DNA in specific
regions of chromosomes across the human genome. This is
termed ______________

A

DNA-based human identification.

67
Q

___________ – not as diverse as those observed
using other systems of identification. This is an older
system of human identification.

A

HLA Polymorphisms

68
Q

__________ – first DNA-based identification system.
Based upon polymorphism in the length of restriction
enzyme-cut products across the genome. It utilizes
Southern blotting and hybridization as a method of
laboratory testing.

A

RFLP Analysis

69
Q

It is most widely used system for identification of
individuals
Ø Very high power of discrimination
Ø ________are typed and compared to the
CODIS database for criminal investigations
Ø _________ typing of older specimens or cold cases
utilizes this system when the specimens are in
acceptable condition
Ø ________uses the same STR system as those
used in the CODIS allele database
Ø _____ engraftment is a clinical application of
this chemistry. STR analysis may be performed to
track donor versus recipient DNA components
following transplant.
Ø Other uses of DNA-based human identification
include QC of _______ specimens to ensure that a
patient matches a specific specimen.

A

STR Analysis

Common alleles 
Forensic DNA
Parentage testing 
Bone marrow
histological
70
Q

____________ – identification of individual
based upon their mitochondrial haplotype.
Ø Primarily used on degraded specimens
Ø Only capable of determining maternally inherited
alleles with this system, because mitochondria are
passed from mothers to all offspring
Ø Often used in cold cases, because there are many
hundreds of copies of mitochondrial DNA present for
every one copy of nuclear DNA. This allows for a
greater possibility of success in analysis.

A

Mitochondrial DNA analysis

71
Q 
\_\_\_\_\_\_\_\_\_\_\_– newer system for human 
identification
Ø Only capable of determining paternally inherited 
alleles
Ø System only used to type males
Ø Based upon repeat length polymorphism
A

Y-chromosome analysis

72
Q

__________– coined by James Watson; referred to the
biology of DNA; study of nucleic acids. Handling and analysis of
nucleic acids, DNA and RNA

A

Molecular Biology

73
Q

DNA

Macromolecule of C, N, O, P, and H atoms; assembled in
units of nucleotides composed of a__________and a __________

Nitrogen bases: adenine, cytosine, guanine, thymine.
They are attached to a deoxyribose sugar which forms a
polymer with the deoxyribose sugars of other
nucleotides through a ______ bond.

A

phosphorylated ribose sugar
nitrogen base.
phosphodiester

74
Q

NUCLEOTIDES

• Each nucleotide consists of a _________ The first
carbon of which is covalently joined to a _____ base
and the fifth carbon to a _____ moiety.
• Can be converted to nucleosides by _______.
• ________: a nitrogen base bound to an
unphosphorylated sugar.
• If the ribose sugar is _______ the molecule is a
nucleoside mono-, di-, or triphosphate or a nucleotide.
• The hydroxyl group on the ____ carbon is important for
forming the _________ that is the backbone
of the DNA strand.
• _______ – double ring structure; _______.
• ________ – single ring structure; ______.
• ________________________are the key to the
specificity of most nucleic acid-based test used in the
molecular laboratory.
• _________________ is also how the
information held in the linear order of the nucleotides is
maintained. As DNA is polymerized, each nucleotide to
be added to the new DNA strand, hydrogen bonds with
the complementary nucleotide on the parental strand
(A:T, G:C)

A 
five-carbon sugar; nitrogen; phosphate
hydrolysis
Nucleoside
phosphorylated,
third; phosphodiester bond
Purines; guanine, adenine
Pyrimidines; thymine, cytosine
Hydrogen bonds between nucleotides 
Specific hydrogen bond formation
75
Q

NUCLEIC ACID

• A macromolecule made of nucleotides bound together
by the _______ and __________ on their sugars.
• A nucleic acid chain grows by the attachment of the _________ group of an incoming nucleotide to the ________ of the last nucleotide on the growing chain.
• Addition of nucleotides in this way gives the DNA chain
a polarity; that is, it has a 5’ phosphate end and a 3’
hydroxyl end. We refer to DNA as oriented in a _________, and the linear sequence of the nucleotides, by
convention, is read in that order.
• The bases are positioned such that the __________
chain that connects them (sugar-phosphate backbone)
is oriented in a spiral or helix around the nitrogen bases.
• The DNA double helix represents two versions of the
information stored in the form of the order or sequence
of the nucleotides on each chain. The sequences of two
strands that form the double helix are ________,
not identical.
• They are in ________orientation, with the 5’ end of
one strand at the 3’ end of the order. Formation of
hydrogen bonds between two complementary strands
of DNA is called ________. Single strands of DNA with
identical sequences will ___ hybridize with each other.

A 
phosphate; hydroxyl groups
5’ phosphate; 3’ hydroxyl group
5’ to 3’ direction
sugar-phosphate
complementary
antiparallel 
hybridization
not
76
Q

DNA REPLICATION

• As DNA synthesis proceeds in the 5’ to 3’ direction, __________, the enzyme responsible for polymerizing the nucleotide chains, uses a guide or template, to determine which nucleotides to add to the chain.
• _____________ is the key to maintaining the sequence of the nucleotides in DNA through new generations. It is important that this information, in the form of the DNA sequence, be transferred faithfully at cell division.
• DNA is first ______ from the helical duplex so that
each single strand may serve as a template for the
addition of nucleotides to the new strand.
• The new strand is then _______ by hydrogen bonding
of the complementary incoming nucleotide to the
nitrogen base on the template strand and then a
nucleophilic attack of the deoxyribose 3’ hydroxyl
oxygen on a phosphorus atom of the phosphate group
on the hydrogen-bonded nucleotide triphosphate.
• _________ is released with the formation of a
phosphodiester bond between the new nucleotide and
the last nucleotide of the growing chain. The duplicated
helix will ultimately consist of one template strand and
one newly synthesized strand.
• The antiparallel nature of duplex DNA and the requirement for the DNA synthesis apparatus to read the template strand in a 3’ to 5’ direction _____ consistent with copying of both strands simultaneously in the same direction.

A 
DNA polymerase
Semi-conservative replication
unwound
elongated
Orthophosphate
are not
77
Q

________– small fragments are the key to explaining how both strands of the parent helix are not copied in the same way.

A

Okazaki fragments

78
Q

__________ – DNA replication proceeds in a

continuous manner on the 3’ to 5’ strand

A

Leading strand

79
Q

__________– 5’ to 3’ strand is copied in a
discontinuous manner from the leading strand’s
replication apparatus which jumps ahead a short
distance on the 5’ to 3’ strand and then copies backward
toward the replication fork.

A

Lagging strand

80
Q

DNA cannot be synthesized _____; a preceding base

must be present to provide they hydroxyl group.

A

de novo

81
Q

______ – RNA synthesizing enzyme that catalyzes the
synthesis of short RNA primers required for priming DNA
synthesis. Must work repeatedly on the lagging strand to
prime synthesis of each Okazaki fragment

A

Primase

82
Q

_____ – unwinds and untangles the double helix at the

replication fork

A

Helicase

83
Q

________ – relaxes the supercoiled DNA

A

Topoisomerase

84
Q

_______– synthesizes the new DNA strand; also

proofreads and corrects some errors

A

Polymerase

85
Q

_______ – rejoins two DNA strands into a double helix and

joins Okazaki fragments of the lagging strand

A

Ligase

86
Q

_________ – used to manipulate DNA in vitro, for instance,
to make stepwise deletions in linearized DNA or to
modify DNA ends after cutting with restriction enzymes

A

Nuclease

87
Q

_______ – catalyzes the addition of methyl
groups to nitrogen bases, usually _____ and ________
in DNA strands

A

Methyltransferase

adenines; cytosines

88
Q

Mispair base removed by _______. DNA polymerase tries a second time

A

exonuclease

89
Q

It is similar to DNA, differs in sugar moieties and in one
nitrogen base component. _____ and _____,
respectively.

• Almost all ______ do not have complementary
partner strands, they are not completely single
stranded. Through __________, RNA species
fold and loop upon themselves to take on a doublestranded character that is important for their function.
RNA can also pair with complementary single strands of
DNA or another RNA and form a double helix.

A

RNA
Ribose; uracil
RNA strands
internal homologies,

90
Q

• The copying of one strand of DNA into RNA by a process
similar to that of DNA replication.
• Activity is catalyzed by _______, which occurs
mostly in _____.

A

Transcription
RNA polymerase
interphase

91
Q

TRANSCRIPTION

_________– synthesis of mRNA, the type of RNA that
carries genetic information to be translate into protein.

A

Polymerase II

92
Q

Transcription _______

Ø Regulated in all organisms so that genes are
transcribed as required by specific cell types.
Ø ________ – where RNA polymerase and its
supporting accessory proteins assemble on DNA
Ø _________ of transcription or RNA synthesis
greatly outnumber DNA initiation sites in both
prokaryotes and eukaryotes.
Ø ______ in prokaryotes and eukaryotes
differ and work with different supporting proteins
to find and bind to DNA in preparation for
transcription.

A

Initiation
Promoter
Initiation sites
RNA polymerases

93
Q

Transcription ______

Ø RNA polymerases in both eukaryotes and prokaryotes synthesize RNA using the base sequence of one strand of the double helix as a guide.
Ø The DNA double helix is locally _____ into single strands to allow the assembly and passage of the transcription machinery, forming a _______.
Ø RNA synthesis does not require _____. Subsequent ribonucleoside triphosphates retain only the _______, the one closest to the ribose sugar.

A

Elongation

unwound
transcription bubble
priming
alpha phosphate,

94
Q

Transcription __________

Ø Accomplished in some genes by interactions
between RNA polymerase and nucleotide signals
in the DNA template.
Ø _____ – helicase enzyme that associates with RNA
polymerase and inactivates the elongation complex at a cytosine-rich termination site in the DNA.
Ø mRNA synthesis is catalyzed by ______ in eukaryotes. It then proceeds along the DNA template until a polyadenylation signal (polyA site) is encountered. At this point, the process of
termination of transcription is activated.
Ø There is _______ sequence in DNA that specifies
termination of transcription.
Ø As the polymerase proceeds pas the polyA site, the nascent mRNA is released by an _________ associated with the carboxy terminal end of polymerase II.
Ø RNA synthesized beyond the site trails out of the
polymerase and is bound by another __________
that begins to degrade the RNA 5’ to 3’ toward the
RNA polymerase.
Ø When the exonuclease catches up with the
polymerase, transcription ______.

A

Termination

Rho
polymerase II
no specific
endonuclease 
exonuclease 
stops
95
Q

Give its template and product of this enzyme:

E.coli RNA polymerase II 
RNA polymerase I
RNA polymerase II
RNA polymerase III
Mitochondrial RNA polymerase
Mammalian DNA polymerase α
HCV RNA polymerase
Dengue virus RNA polymerase
PolyA polymerase
A 
DNA - mRNA
DNA - rRNA
DNA - mRNA
DNA - tRNA, snRNA
DNA - mRNA
DNA - primers
RNA - viral genome
RNA - viral genome
none - PolyA tails
96
Q

Types and Structures of RNA:

Ø Largest component of cellular RNA, comprising 80-
90% of the total cellular RNA. An important structural
and functional part of the _____, cellular
organelles where proteins are synthesized.

A 
Ribosomal RNA (rRNA)
ribosomes
97
Q

Ø Initial connection between information stored in DNA
and translation apparatus that will ultimately produce
the protein products responsible for the phenotype.
Ø Amount of particular mRNA is related to the
requirement for its final product.

A

Messenger RNA (mRNA)

98
Q

________– some messages are transcribed constantly and are relatively abundant in the cell

A

Constitutive transcription

99
Q

__________– transcribed only at certain times during the cell cycle or under particular conditions

A

Inducible or regulatory transcription

100
Q

__________ – most messengers carry a sequence of polyadenylic acid at the _____, a polyA tail. It is not coded in genomic DNA, but added to the RNA after synthesis of the pre-mRNA.

A

Polyadenylation

3’ terminus

101
Q

____________ – enzyme responsible for

adding the adenines to the end of the transcript.

A

Polyadenylate polymerase

102
Q

__________– eukaryotic mRNA is blocked at the ______ by a 5’-5’ pyrophosphate bridge to a methylated guanosine. The structure is called a cap. It confers a protective function and serves as a recognition signal for the translational apparatus.

A

Capping

5’ terminus

103
Q

________ – prokaryotic structural genes contain
____ lengths of open reading frame, sequences that code for amino acids. In eukaryotic coding regions, they are interrupted with long stretches of noncoding DNA sequences called _____.
They are then spliced or removed, and the remaining
sequences that code for the protein product are ____.

A

Splicing

uninterrupted
introns
exons

104
Q

_________ – modifies products of genes by alternate insertion of different exons.

A

Alternative splicing

105
Q

Ø Functions in splicing in eukaryotes
Ø Stays in nucleus after its transcription by RNA
polymerase I or III
Ø Isolated from _____ and cervical carcinoma cell
lines

A

Small nuclear RNA (snRNA)

hepatoma

106
Q

Ø Relatively short, single-stranded polynucleotides.
There is at least one ___ for each amino acid.
Ø Translation of information from nucleic acid to
protein requires reading of the mRNA by ______,
using adaptor molecules or tRNA.
Ø Variable loop is larger in longer tRNAs. Another
seven-base loop is larger in longer tRNAs

A

Transfer RNA

ribosomes

107
Q

RNA POLYMERASES

• RNA synthesis is catalyzed by _________enzymes.
• One ___________ enzyme is responsible for
the synthesis of all types of RNA in the prokaryotic cell.
• Eukaryotes have three different RNA polymerase
enzymes. All of these enzymes are __________ RNA
polymerases; that is, they require a DNA template.
• __________RNA polymerases require and RNA
template.

A

RNA polymerase
multisubunit prokaryotic
DNA-dependent
RNA-dependent

108
Q

__________ – degrade RNA in a manner similar to the
degradation of DNA by ________. They are
ubiquitous and stable that degrade all types of RNA. RNA
work is performed because they are very resistant to
inactivation.

A

Ribonucleases

deoxyribonucleases

109
Q

__________– RNA synthesis and processing require the

activity of helicases to catalyze the unwinding of doublestranded RNA. Some of them work exclusively on RNA.

A

RNA Helicases

110
Q

Ø Products of transcription and translation of nucleic
acids.
Ø Polymers of amino acids. Each amino acid has
characteristic biochemical properties determined by
the nature of its amino acid side chain.
Ø The most abundant macromolecules in cells. The
collection of proteins encoded in all of an organism’s
DNA is a _____.
Ø The ultimate effect of the information stored and
delivered by the nucleic acid is manifested in
____.

A

Proteins

proteome
proteins

111
Q

Ø Even if proteins are not being tested directly, they
manifest the _______ directed by the nucleic acid
information

A

phenotype

112
Q

In order to interpret the results of nucleic acid
analysis accurately, it is important to understand the
______________ from DNA to
protein as dictated by the genetic code.

A

movement of genetic information

113
Q

__________

Ø Grouped according to their _____ or tendency to
interact with water at ___.
Ø _____, ________ polar, _______ charged
polar, and ______ charged polar.
Ø Properties of amino acids that make up a protein
determine the ___ and biochemical nature of the
protein. A single protein can have separate domains
with different proteins.
Ø Synthesized in vivo by ______ enzymes so that
naturally occurring proteins are made of amino acids
of L-stereochemistry.
Ø The ionization can switch between the amino and
carboxyl groups, making the amino acids _______
at physiological pH.

A

Amino acids

polarity; pH 7
Nonpolar, uncharged, negatively, positively
shape
stereospecific
zwitterions
114
Q

Primary structure

§ The sequence of amino acids in a protein
determines the nature and activity of that
protein.
§ Read by convention from the _________ to the _________end.
§ _____ changes in primary structure can alter
the activity of proteins dramatically, because
the amino acids must _____________to bring about protein structure and function.

A

amino terminal end; carboxy terminal
Minor
often cooperate with one another

115
Q

Ø Secondary structure
§ Interactions between amino acid side chains
fold a protein into _________.
§ Some proteins, especially structural proteins,
consist almost entirely of _________
§ Globular proteins have varying amounts of
alpha helices and beta sheets.
§ These include ordered beta or beta-pleated
sheets and less-ordered alpha helices, or
random coils; alpha-helix and beta-sheet
structures in proteins.

A

predictable configurations

alpha helices or beta sheets.

116
Q

Tertiary structure
§ Secondary structures are further folded and
arranged into a ___________.
§ If a protein loses its tertiary structure, it is
______.
§ ______ in DNA that substitute different
amino acids in the primary structure can also
alter tertiary structure.

A

tertiary structure
denatured
Mutations

117
Q

§ ________ – proteins denatured by heat or by
conformations forced on innocuous peptides.
Aggregations of prions-induced folded
proteins cause transmissible __________, such as ________and bovine spongiform encephalitis
or _______.

A

Prions

spongiform encephalopathies

Creutzfeldt-Jakob disease
mad cow disease

118
Q

Quaternary structure
§ Combinatorial nature of protein function may
account for the __________ of higher
organisms without a concurrent increase in
gene number.
§ Monomer, dimer, trimer, tetramer,
oligomers.

A

genetic complexity

119
Q

_________
Ø Ordered sequence of nucleotides on a chromosome
Ø Encodes a specific functional product
Ø Fundamental ____and functional unit of
inheritance
Ø First studied by tracking mutations that took away
their function and observing the resulting phenotype
Ø Part of a chromosome responsible for the phenotype
affected by _____
Ø Contains structural and regulatory sequences
important for the regulated expression of the gene

A

Gene

physical
mutation

120
Q

Ø ___________ – dictionary to translate the 4-
nucleotide sequence information in DNA to the 20-
amino acid sequence information in proteins

A

Genetic Code

121
Q

TRANSLATION

After transcription of the sequence information in DNA to
RNA, the transcribed sequence must be transferred into
_______
• Through the genetic code, a specific nucleic acid sequence
is translated to an amino acid sequence and, ultimately, to
a ______.
• Takes place on ____, __________ particles
first observed by electron microscopy of animal cells.

A

proteins.
phenotype
ribosomes
ribonucleoprotein

122
Q

_______ – molecular factor that can recognize components of

both nucleic acid and protein sequences.

A

tRNA

123
Q

TRANSLATION

In contrast to the analysis of phenotypic traits
(microscopic and colonial morphologies, enzyme or
pigment production, carbohydrate fermentation
patterns), the analyte for molecular testing is the _______,
transcriptome, or ______ of the microorganism.

A

genome

proteome

124
Q

Bacteria, fungi, and parasites have _____ genomes,
whereas viruses can have ______genomes. Prions,
which cause transmissible encephalopathies such as
Creutzfeldt-Jakob disease, consist only of _________

A

DNA
DNA or RNA
protein

MOLBIOF!NA1S (9 decks)

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