Topic 1C Flashcards

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1
Q

what are key elements of info needed to understand DNA replication?

A

strands are antiparallel, purine base pairs with pyrimidine base

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2
Q

template strand

A

in DNA replication, the parental strand whose sequence is used to synthesize a complementary daughter strand

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3
Q

daughter strand

A

in DNA replication, the strand synthesized from a parental template strand

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4
Q

semiconservative replication

A

the mechanism of DNA replication in which each strand of a parental DNA duplex serves as a template for the synthesis of a new daughter strand

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5
Q

after replication, what does each new DNA duplex consist of?

A

one strand that was originally part of the parental duplex and one newly synthesized strand

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6
Q

what does conservative replication (the alternative model) propose?

A

the original DNA duplex remains intact and the daughter DNA duplex is completely new

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7
Q

who’s research and experimental evidence supported semiconservative replication?

A

Meselson and Stahl-used two non-radioactive isotopes of nitrogen and measured density of DNA

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8
Q

replication fork

A

the site where the parental DNA strands separate as the DNA duplex unwinds

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9
Q

new DNA strands grows in what direction?

A

can only grow by adding successive nucleotides to the 3’end-grows in a 5’ to 3’ direction

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10
Q

DNA polymerase

A

an enzyme that is a critical component of a large protein complex that carries out DNA replication (highly conserved)- synthesizes a new DNA strand from an existing template*

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11
Q

what can correct mistakes in replication?

A

DNA polymerase

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12
Q

leading strand

A

a daughter strand that has its 3’ end pointed toward the replication fork, so as the parental double helix unwinds, this daughter strand can be synthesized as one long, continuous polymer

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13
Q

lagging strand

A

a daughter strand that has its 5’ end pointed toward the replication fork, so as the parental double helix unwinds, a new DNA piece is initiated at intervals, and each new piece is elongated at its 3’ end until it reaches the piece in front of it

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14
Q

Okazaki fragment

A

in DNA replication, any of the many short DNA pieces in the lagging strand

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15
Q

what explains the presence of leading and lagging strands during DNA replication?

A

the antiparallel nature of the two strands in a DNA double helix and the fact that DNA polymerase can synthesize DNA in only one direction

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16
Q

primer

A

a short stretch of RNA at the beginning of each new DNA strand that serves as a starter for DNA synthesis

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17
Q

why is a primer needed?

A

because DNA polymerase cannot begin a new strand on its own, can only elongate the end of an existing piece of DNA or RNA

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18
Q

RNA primase

A

an RNA polymerase that synthesizes a short piece of RNA complementary to the DNA template and does not require a primer

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19
Q

where is the section of RNA primer located?

A

at the 5’ end, the lagging strand has many primers for each fragment

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20
Q

in the lagging strand, when a growing fragment comes into contact with the primer, what happens?

A

a different DNA polymerase removes RNA primer and extends fragment of DNA to fill space left over. DNA ligase joins fragments

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21
Q

DNA ligase

A

an enzyme that uses the energy in ATP to close a nick in a DNA strand, joining the 3’ hydroxyl of one end to the ‘ phosphate of the other end

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22
Q

topoisomerase II

A

an enzyme that breaks a DNA helix, rotates the ends and seals the break-relieves stress on the double helix that results from its unwinding at the replication fork

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23
Q

helicase

A

a protein that unwinds the parental double helix at the replication fork

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24
Q

single-strand binding protein

A

a protein that binds single-stranded nucleic acids-prevents template strands from coming back together

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25
Q

how does synthesis of DNA strands occur at the same time and rate?

A

the synthesis is coordinated because: polymerase complexes for each strand stay in contact with each other-polymerase passes through in the same direction, lagging strand is looped until new legging strand encounters previous fragment: 3’ end of both strands are elongated together

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26
Q

what happens to rate of replication when DNA damage occurs?

A

rate of synthesis slows down so that DNA can be repaired

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27
Q

if synthesis of one strand slows down to repair DNA damage, what happens to the other strand?

A

the synthesis slows down too

28
Q

when is pairing of the replication complexes disrupted?

A

when the RNA primer of the previous Okazaki fragment is encountered and when new lagging-strand primer is formed

29
Q

proofreading

A

the process in which DNA polymerases can immediately correct their own errors by excising and replacing a mismatched base

30
Q

origin of replication

A

any point on a DNA molecule at which DNA synthesis is initiated

31
Q

replication bubble

A

a region formed by the opening of a DNA duplex at an origin of replication, which has a replication fork at each end

32
Q

while prokaryotic chromosomes only have …

A

one origin of replication, eukaryotic chromosomes have many

33
Q

each replication bubble has:

A

two replication forks that move in opposite direction

34
Q

within a single replication bubble, the same daughter strand is:

A

a leading strand at one replication fork and the lagging strand at the other replication fork

35
Q

circular DNA molecules typically have:

A

only one origin of replication

36
Q

circular DNA molecules can be replicated completely because:

A

it has no ends, replication forks move completely around the circle

37
Q

linear DNA molecules have ends, and after each round of DNA replication….

A

the ends become slightly shorter

38
Q

where is a primer required in the leading strand?

A

only at the origin of replication

39
Q

what is the purpose of the final RNA primer in the lagging strand?

A

this primer initiates synthesis of the final Okazaki fragment of the lagging strand, however this means that there is no other Okazaki fragment to synthesize the missing 100 base pairs and remove the primer

40
Q

telomere

A

a repeating sequence at each end of a eukaryotic chromosome (attaches to template strand-3’ end, allows formation of original telomere sequence in daughter strand)

41
Q

telomerase

A

an enzyme containing an RNA template from which complementary telomere repeats are synthesized

42
Q

once the 3’ end of the template strand has been elongated by the telomere, …

A

an RNA primer and the complementary DNA strands are synthesized to restore the original telomere which can synthesize another length of the original telomere in daughter strand

43
Q

the slight shortening and subsequent restoration that takes place has no harmful consequences because…

A

telomeres have no genes

44
Q

germ cells

A

the reproductive cells that produce sperm or eggs and the cells that give rise to them

45
Q

stem cell

A

an undifferentiated cell that can undergo an unlimited number of mitotic divisions and differentiate into any of a large number of specialized cell types

46
Q

where is telomerase activity most active?

A

in germ cells and stem cells

47
Q

describe telomerase activity in most cells in the adult body:

A

inactive-telomere is actually shortened 100 base pairs in each mitotic division which limits the number of mitotic divisions that cells can undergo

48
Q

human cells stop dividing when:

A

their chromosomes have telomeres with fewer than about 100 copies of the telomere repeat

49
Q

what is the telomere repeat sequence?

A

3’-GGGATT-5’

50
Q

polymerase chain reaction (PCR)

A

a selective and highly sensitive method for making copies of a piece of DNA, which allows a targeted region of a DNA molecule to be replicated into as many copies as desired

51
Q

amplified

A

in PCR technology, an alternative term for replicated

52
Q

when is PCR used? (hint: PCR is both selective and highly sensitive)

A

it can be used to amplify and detect small quantities of nucleic acids

53
Q

what does the PCR solution require:

A

template DNA (including region to be amplified), DNA polymerase, all nucleotides, primers (amount greater than number of template DNA molecules)

54
Q

oligonucleotide

A

a short (typically 20 to 30 nucleotides), single-stranded molecule of known sequence produced by chemical synthesis; oligonucleotides are often used as primer sequences in the polymerase chain reaction

55
Q

the base sequences of oligonucleotides are chosen to be:

A

complementary to the ends of the region of DNA to be amplified

56
Q

what are the three steps of PCR?

A
  1. denaturation
  2. annealing
  3. extension
57
Q

denaturation

A

the unfolding of proteins by chemical treatment or high temperature; the separation of paired, complementary strands of nucleic acid (breaks H-bonds*)

58
Q

annealing

A

the coming together of complementary strands of single-stranded nucleic acids by base pairing-primers anneal to complementary sequence on the DNA

59
Q

extension (PCR)

A

a step in the PCR for producing new DNA fragments in which the reaction mixture is heated to the optimal temperature for DNA polymerase, and each primer is elongated by means of deoxynucleoside triphosphates

60
Q

PCR is repeated over and over (25-35 cycles) and the number of copies of the targeted fragment…

A

doubles (2^n), after third round of amplification, only molecules as long as the region of the template that is complementary to the primers is produced

61
Q

why do you have to continue adding fresh DNA polymerase?

A

because the enzyme loses structure and function at the high temperatures during the denaturation stage

62
Q

in modern times, DNA polymerase from what bacteria is used?

A

Taz polymerase (resistant to heat, lived in hot springs)

63
Q

how do you determine whether or not PCR has yielded the expected product?

A

by determining the size of the amplified DNA molecules (compares expected size to actual size-by gel electrophoresis)

64
Q

gel electrophoresis

A

a procedure to determine the size of a DNA fragment, in which DNA samples are inserted into slots or wells in a gel and a current is passed through. Fragments move toward the positive pole according to size

65
Q

in a sample of gel electrophoresis, where are the larger fragments located?

A

near the top, moves less in a given interval of time compared to shorter fragments