LEC12 - DNA Replication DNA Replication (S phase) Cell...

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DNA Replication DNA Replication (S phase) Cell Cell Division Division (Mitosis) (Mitosis)
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New strand Old strand Old strand Models for DNA replication (Meselson-Stahl, 1958) Conservative Semi-conservative
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How can one distinguish between the original strands of DNA and the newly synthesized strands? Density label DNA, separate with equilibrium centrifugation on a CsCl gradient
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Experiment: Grow cells in media containing 15 N Shift to media containing 14 N Extract DNA and centrifuge after 1 and 2 generations Predictions Conservative Semi-conservative HH HH H L H L HH LL 1st doubling in 14 N
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HH LL H L H L H L H L LL LL HH LL LL LL 2nd doubling in 14 N
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Visualization of semi- conservative replication using BrdU in eukaryotes •Combination of fluorescent dye and Giemsa stain distinguishes between chromosomes with one strand containing BrdU and those with two strands of BrdU Br Two strands containing BrdU (light) One strand containing BrdU (dark)
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DNA synthesis initiates at a single site in prokaryotes: Origin of Replication In eukaryotes, there are many origins of replication per chromosome
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This note was uploaded on 07/08/2009 for the course LIFESCI LS 3 taught by Professor Paulolague during the Fall '09 term at UCLA.

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LEC12 - DNA Replication DNA Replication (S phase) Cell...

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