Handout for Lecture 2 - C3032

 

Information You Should Know: DNA to Protein

I. Books for enrichment and background
A. Double Helix - James Watson
B. Girls, Genes, and Gamow - James Watson
C. Eighth Day of Creation - Horace Freeland Judson
D. What Mad Pursuit - Francis Crick
E. Molecular Genetics, 2nd edition - Stent and Calendar

II. Experiments demonstrating the importance of DNA (but also that genetic material can be transferred chemically)
A. Griffiths (1928): the demonstration of transformation
B. Avery, MacLeod, and McCarty (1944) the experiment that should have won a Nobel prize: the isolation of the transforming activity.
C. Hershey and Chase and the Waring Blender Experiment

III. Watson Crick Model (1953)
A. Basic structure
B. Model: complementarity through antiparallel strands
C. The angle that the sugars form with the bases gives rise to a larger space (the major groove) and a smaller space (minor groove) along the helix
D. Strands are antiparallel
E. 5' vs. 3'

IV. Meselson/Stahl Experiment - Support for semiconservative replication
A. Made heavy molecules in E. coli with 15N and then switched bacteria to media with 14N
B. Isolated DNA and used CsCl centrifugation to separate molecules of different density.

V. Transcription (DNA -> RNA)
A. Differences between RNA and DNA
B. Information to review
C. RNA polymerase in E. coli (not used to make primer) complete enzyme (holoenzyme) 480 kd is made of
D. in vitro transcription systems are available for both prokarotes and eukaryotes and have permitted the identification of components needed for transcription and its control.
E. Eukaryotes RNA and polymerases

VI. Translation
A. Components
B. Direction of synthesis: N to C.
C. In vitro translation systems are available. The most common eukaryotic systems are from wheat germ and rabbit reticulocytes.