Bio. 551 Rec. DNA

Dr. Michelle Mardahl

Exam II. Study Sheet 40 points:

Topics:


In Vitro Mutagenesis- Reverse Genetics- 25 points

Gene Therapy- 5 points

Transgenic Eggs & Animals- 10 points

 

 

Be prepared to use the following to design an experiment to mutate a gene in vitro:

Elimination of restriction site

Linker insertion mutagenesis

Nested Deletions

Dut-ung

Dpn I-

Taq DNA ligase

Cassette Mutagenesis

What are the parameters for primer design for in vitro mutagenesis?

How are mutants screened for using PCR, SSCP?

 

Terminology- Gene Therapy Approaches

Ex vivo

In situ

In vivo

Chimeraplasty -

Compare & Contrast the advantages & disadvantages of different gene therapy vectors including

a. liposomal

b. viral

c. conjugated DNA

d. transpon-mediated

Compare & contrast the different viral vectors currently in use.

What are scientific & ethical concerns about gene therapy?

Diagram how to make and screen for:

a transgenic fly

transgenic mouse

knock-out mouse

conditional knock-out mouse