Study Guide to Exam IV, Microbial Genetics

 

Vocabulary

Replicon
Copy number
Par
Hok/Sok
Incompatibility
Curing
Host restriction phenomenon
hsdMRS
Arber
Smith/Nathans
restriction endonuclease
HindII
Type II restriction endonucleases
EcoRI
sticky ends
physical mapping
Cohen and Boyer
Berg/homopolymeric tailing
cloning vectors
blue-white selection
polylinker (multiple cloning site)
genomic DNA library
insertional inactivation
functional complementation
colony hybridization
plaque hybridization
Southern blot (Northern, Western, etc)
immunological screening

 

 

Questions and problems.

A circular DNA molecule is digested with EcoR1, HindIII, PstI, and three double digests.  Fragment sizes are as follows: EcoR1, 8 kb; HindIII, 7 kb and 1 kb; EcoR1/HindIII, 5kb, 2 kb, and 1 kb; Pst, 8 kb; EcoR1/Pst, 6 kb and 2 kb; Pst/HindIII, 4 kb, 3 kb, and 1 kb.  Map the sites.

 

 

What are the general steps involved in cloning a gene?  What are some methods for identifying a particular clone?

 

 

Which of the following are more likely to be a Type II RE recognition site? (only one strand is shown)

GAATCG,  GATATC,  AAATTT,  ACGGCA