DNA replication and manipulation Flashcards

1
Q

Where does DNA replication occur

A

S phase

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2
Q

What shape is DNA in

A

helix w two antiparallel strands

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3
Q

Why do hydrogen bonding inbetween DNA strands allow it to unravel

A

bc they are weaker

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4
Q

What direction is DNA made

A

5’-3’ only

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5
Q

DNA replication conservative model

A

no mixing DNA

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6
Q

Semiconservative model

A

evenly split mixed DNA

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7
Q

Dispersive model

A

random mixing DNA

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8
Q

Which experiment showed that DNA replication is semi-conservative

A

messelsohn and stahl

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9
Q

How did messelsohn and stahl do their experiment

A

growing bacteria in lighter and lighter nitrogens and DNA forms bands when centrifuged

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10
Q

Draw out semiconservative model

A

Dark strand on first, split half on second, and light and half on third

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11
Q

If DNA replication was conservative which bands would be observed after one round of replication

A

heavy and light

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12
Q

Complications in DNA replication(4)

A
  1. unwinding tension
  2. only add nucleotides to existing nucleotide
  3. opposite directions on each strand
  4. pieces need to be joined together
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13
Q

Helicase

A

unwinds double helix at replication forks

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14
Q

Single-strand binding protein

A

binds and stabilizes single stranded DNA when it is used as a template

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15
Q

topoisomerase

A

relieves strain of overwinding by breaking, swiveling, and rejoining DNA strands

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16
Q

primase

A

makes RNA primer at 5’ of leading and 5’ of each okazaki fragment of laggiing

17
Q

DNA pol III

A

uses parental DNA as template and makes new DNA by adding nucleotides to RNA primer

18
Q

DNA pol I

A

removes RNA primers and replaces with DNA nucleotides

19
Q

DNA ligase

A

joins okazaki fragments on lagging strand
leading strand: joins 3’ to rest of leading

20
Q

Step 1 of replication (3)

A
  1. helicase unwinds
  2. topoisomerase relieves stress
  3. single-stranded binding proteins stabilize single strands
21
Q

Step 2 of replication

A
  1. RNA primer attaches to 5’ primer
  2. elongation as bases are added
  3. RNA primers are replaced with DNA
  4. pieces in the lagging strand and put together
22
Q

Prokaryote replication

A

single origin and moves around the circular c chromosome in both directions

23
Q

Difference between eukaryotic and prokaryotic replication

A

prokaryotic one origin
eukaryotic many origins of replication

24
Q

Telomeres

A

protect ends of chromosomes

25
Relation between age and telomere length
As age increases, telomere length decreases
26
End replication problem
few bases at '5 end of new strands are missing
27
Telomerase
some cells express this carries its own RNA template
28
Some types of cancer cells express telomerase to
divide a greater number of times before dying
29
Polymerase Chain Reaction purpose
make many copies of a DNA fragment in vitro
30
Primers
oglionucleotides
31
Things you need in a PCR(5)
1. DNA template 2. Primers 3. DNA polymerase 4. dNTPs 5. Buffer/cofactors
32
Restriction enzymes
recognize and cut specific sequences of DNA
33
EcoRI restriction endonuclease
restriction enzyme
34
restriction enzymes with plasmids
used to insert genes/DNA fragments of interest into a vector
35
agarose gel electrophoresis
used to visualize DNA
36
shorter strands
migrates faster
37
larger strands
migrate slower
38
larger strands
migrate slower
39
CRISPR
used to insert and change gene sequences