DNA replication Flashcards
whats semi conservative DNA
when replicated, dsDNA = 2x ( parent + new antiparallel strand )
whats conservative DNA
when replicated, one dsDNA is the same, and one is completely new - synthesised complementarily to parent
whats dispersive DNA
daughter DNA contains parental and newly synthesised DNA
describe Meselson Stahl experiment
e coli grown in 15N mediul
this was added to 14N medium, so any newly synthesised DNA would contain 14N
separated in centrifuge - more dense (15N) at bottom, less dense at top.
what is the position where the DNA helix is unwinded called
replication origin
how many replication origins do cprokaryotic and eukaryotic chromosomes have?
1, multiple
describe regulation of prokaryotic origin of replication
highly regulated - only opened when A’s are methylated and when sufficient nutrients are available
describe first steps of dna rep (hint: helicase and ssBP)
- DNA helicase opens DNA helix
• DNA helicase hydrolyses ATP; harnesses this energy to pry apart 2 strands. - ssBP keeps strands separate, but keeps bases exposed
describe what happens after ssBP but before DNA p
- DNA primase synthesises RNA primer
- Sliding clamp and clamp loader bind together
- complex binds to RNA primer
- DNA p binds to primer and complex
- Clamp loader dissociates
describe how DNA p works
- DNAp reads the template strand in 3’5’ direction
- DNAp forms 5’3’ complementary DNA strand
• DNAp has fingers, which recognise the other strand’s base, and adds the complementary base.
• DNAp hydrolyses nucleoside triphosphates into nucleoside monophosphates – first pyrophosphate is formed, then 2x phosphate groups
• DNAp catalyses condensation between 3’ OH and OH of 5’ phosphate
what happens to RNA primer when polymer is synthesised
RNase H degrades it. DNA replaces the primer
describe the action of DNA polymerase.
shaped like a hand - groove determines what complementary dNTP to add.
enzyme hydrolyses dnTP to nucleotides and pyrophosphate, enabling addition to the 3’ end.
the half of the strand closest to the 5’ end of template is called the
leading strand
the half of the strand closest tothe 3’ end of the template strand is called the
lagging strand
whats special about lagging strand?
grows via okazaki fragments
they are separated, and joined by ligase