10/13 Flashcards
What are the four tasks/obstacles of DNA replication?
Tasks/obstacles:
1) a lot of genetic material/cell
2) lots of cell divisions
3) must be extremely accurate (= fidelity) so as not to accumulate mutations or lose genetic info
4) process must occur rapidly
What do the complementary nature of the strands in the double helix suggest?
This suggests that each strand could serve as a template for synthesis of a new complementary/matching strand.
Who proposed the semi conservative model of DNA replication?
Watson and Crick
Semi conservative model of DNA replication:
1)
2)
1) each parental strand of dbl helix acts as a template to assemble a NEW daughter strand of DNA that is complementary to itself
2) the result of each new molecule of DNA would consist of one parental strand and one new daughter strand
* **parental DNA is semi conserved in new molecule = (1/2)parent and (1/2)new
Conservative model of DNA replication:
Original parent. DNA is fully conserved in that the entire molecule of parental DNA serves as a template for entirely new molecule composed of only newly synthesized daughter DNA
Dispersive model of DNA replication:
Original double helix is broken down into manageable fragments. Each fragment serves as a template to make new daughter DNA and ends w reassembly of fragments into two complete molecules. (Dispersive pattern of old and new.)
____________ and _______ conducted an experiment to determine which replication model was correct.
Messelson and Stahl conducted an experiment to determine which replication model was correct.
Messelson and Stahl needed a way to distinguish between old and new DNA. They used:
They used heavy N15 and light N14 isotopes to label the nitrogenous bases of DNA. This allowed them to distinguish DNA by their densities.
What three possible arrangements could Messelson and stahl get?
14/14
14/15
15/15 (heaviest)
M&S used ______ density gradient to “band” out molecules based on their densities.
M&S used CsCl2 density gradient to “band” out molecules based on their densities.
What were the three main steps in the M&S experiment?
1) they grew E. Coli for several generations in the presence of N15 (aka all strands of DNA are marked w N15)
2) switched cells to grow on light N14
3) they analyzed the DNA after one and two rounds of sequential replication
What did M&S expect to see if the replication model was semi conservative?
They expected to see half 14/15 molecules and half 15/15 molecules.
What did M&S expect to see if the replication model was conservative?
They expected to see 25% 15/15 molecules and 75% 14/14 molecules.
What did M&S expect to see if the replication model was dispersive?
They expected to see 100% 14/15 molecules.
Results of M&S supported a _________________ model of replication.
Results of M&S supported a semi conservative model of replication.
Replicon
Replicon: an individual unit of replication
Replication origin
Replication origin: point on a xsome where replication initiates
Each replicon contains a ____________ of replication.
Each replicon contains a single origin of replication.
Replication bubble
Replication bubble: the unwinding of dbl helix centered over the replication origin
Replication fork:
Replication fork: active site of DNA replication as it moves down the molecule
Theta replication:
Theta replication: common to circular genomes. EX: bacteria, xsome generates a replication intermediate that resembles Greek letter theta. First described by John Cains
Cains experiment (E. Coli):
1)
2)
3)
1) grew bacteria in presence of tritiated-thymidine (radio label for thymine bases)
2) sampled cells at various time points during replication and each sample of cells were lysed open
3) exposed slide to x-Ray film, which gave him a snapshot of the replication process
Conclusion of Cains experiment:
1)
2)
1) E. Coli xsome has a single origin of replication
2) DNA unwinds at origin (OriC) and generates localized area of single stranded DNA that can serve as a template to make a new daughter DNA
Replication of the E. Coli xsome is ______________.
Replication of the E. Coli xsome is bidirectional.