DNA Replication

Question 0

the newly copied DNA (reverse compliment)

Answer 0

daughter strands

Question 1

each strand is used as a template from which new DNA is copied

Answer 1

template strands

Question 2

where DNA replication begins

Answer 2

origin of replication

Question 3

______________ of replication encoded by specific sequence (oriC)

Answer 3

initiation

Question 4

__________ bind to dnaA box sequences

-5 recognition sites in oriC

Answer 4

proteins

Question 5

_________ bind to each other

Answer 5

proteins

Question 6

DnaC proteins recruit ____________

Answer 6

helicases

Question 7

________ hydrolysis fuels helicases

Answer 7

ATP

Question 8

helicases break __________ bonds

Answer 8

hydrogen

Question 9

uncoils DNA ahead of the Helicases

Answer 9

DNA gyrase (aka topoisomerase II)

Question 10

single stranded binding proteins

-prevent strands from re-annealing

Answer 10

single stranded binding proteins

Question 11

synthesizes short strands of RNA (primers)

Answer 11

primase

Question 12

start the process of DNA replication

Answer 12

RNA primers

Question 13

leading and lagging strands:
__________: unzipped 3’-5’, one RNA primer
__________: unzipped 5’-3’, several RNA primers

Answer 13

leading, lagging

Question 14

catalyzes covalent bond formation between nucleotides

Answer 14

DNA polymerase III

Question 15

• cannot begin DNA synthesis without primers
• can only build new strands 5’-3’ (moving 3’-5’ along template DNA)
• attaches dNTPs to 3’ end of primer (dNTP:deoxiribonucleoside triphosphate, aka nucleosides)

Answer 15

DNA polymerase III

Question 16

one leading strand synthesized

Answer 16

advancing the replication fork

Question 17

Dna fragments constituting the lagging strand

Answer 17

okazaki fragments

Question 18

completion of lagging strand synthesis:

Answer 18

• remove RNA primers (dna polymerase I)
• synthesize DNA in place of RNA primers (dna polymerase I)
• covalently bond adjacent dna fragments (DNA ligase)

Question 19

catalyzes formation of covalent bond between adjacent okazaki fragments.

Answer 19

DNA ligase

Question 20

ter sequences (elements)

Answer 20

termination sequences

Question 21

substance protein Tus

-binds to recognition sequences

Answer 21

termination utilization

Question 22

when replication forks meets Tus, replication _______

Answer 22

ends

Question 23

helicase, primase, and the polymerases are physically connected to each other

Answer 23

replisome

Question 24

AT/GC pairing is ___________ | -1 error in 1000 nucleotides replicated

Answer 24

stable

Question 25

shape/structure of DNA polymerase: - correct pairing causes conformational change in DNA _____________, allowing catalysis of nucleotide incorporation. - 1 error in 100k to 1 mil nucleotides

Answer 25

polymerase

Question 26

similarities of eukaryotic/pro DNA replication

Answer 26

- enzymes (helicase, gyrase, SS binding proteins, primase, plymerase, ligase) - not as well understood as bacterial bc its more complex (large linear chromosomes, chromatin tightly packed, cell cycle regulation more complicated)

Question 27

eukaryotic DNA replication has __________ origins of replication -evidence (1968) ??

Answer 27

multiple | pulse radiolabeled dNTPs to dividing cells

Question 28

more than _____ types of DNA polymerase in mammals

Answer 28

12

Question 29

RNA primer construction:

Answer 29

primase + alpha polymerase | - 10 bp RNA followed by 20-30 bp DNA

Question 30

leading strand synthesis

Answer 30

polymerase epsilon

Question 31

lagging strand synthesis

Answer 31

polymerase sigma

Question 32

several polymerases repair _______ | -polymerase ______-removed incorrect bases

Answer 32

DNA | beta

Question 33

lesion-replicating polymerases -act where aberration occur in DNA _____________ ___________ ______________

Answer 33

abnormal bases | crosslinks

Question 34

removes DNA primer

Answer 34

flap endonuclease

Question 35

replicating the ends of the chromosomes

Answer 35

telomeres

Question 36

telomeres prevent ____________ shortening. | discovered in 1984 ___________&___________

Answer 36

chromosome | Greider & Blackburn

Question 37

telomerase relationship to cancer treatment and aging:

Answer 37

- telomerase inhibitors can stop cell division | - Active telomerase can prevent senescence in cells

Question 38

- template DNA - primers (forward and reverse) - dNTPs - polymerase Taq (originally isolated from bacteria in geothermal pools in yellowstone park) - proper chemical conditions

Answer 38

PCR ingredients

Question 39

PCR cycle:

Answer 39

1. denature DNA at 95 degrees C (apply heat to break hydrogen bonds & make DNA single stranded) 2. anneal DNA primers at 40C-65C (primers brackets fragment to be amplified) 3. extend DNA copies at 72C (optimal temp. for polymerase) 4. denature DNA at 95C 5. anneal DNA primers at 40C - 65C 6. extend DNA copies at 72C 7. cycling is run up to 45 times 8. results in >1,000,000,000 copies of target template DNA

Question 40

used to amplify RNA sequence with DNA (RNA is unstable compared to DNA) - produces cDNA (c= complimentary) - cDNA can be subjected to PCR

Answer 40

reverse transcriptase PCR

Question 41

ingredients for reverse transcriptase PCR

Answer 41

primers reverse trandcriptase dNTPs

Question 42

reverse transcriptase PCR denatures at ______

Answer 42

65C

Question 43

incubate at _______-_______ to transcribe RNA to cDNA in reverse transcriptase PCR

Answer 43

24-37C