Recombinant DNA

Question 1

What is a genomic library?

Answer 1

Cloned colonies with specific restriction fragments

Question 2

What is a cDNA (expression) library?

Answer 2

Cloned colonies with specific gene sequence

Question 3

What is a palindrome sequence? Use?

Answer 3

4 to 8 bp with inverted repeats, serve as site of recognition of restriction endonucleases

Question 4

What are the problems of gene delivery vectors on gene therapy associated with retroviruses?

Answer 4

• host cell replicate successful infection
• random integration of retroviral genome ➡️ risk integration near and activate oncogene ▶️ in X-linked SCID Leukemia reported

Question 5

What is the advantage and disadvantage of use adenovirus as a gene delivery vector?

Answer 5

• advantage: can invade cells that are not dividing

- disadvantage: adenovirus DNA is not integrated into the host cell’s chromosome and is eventually lost

Question 6

What is the most common use of transgenic”knock in” mice?

Answer 6

Studying dominant gene effects, not good for recessive disease

Question 7

What is the advantage of transgenic mice produced introducing the transgene by embryonic stem cells?

Answer 7

• select the cells with the transgene in the cultures
• create knock in (transgene) or knock out (delete) mice (homozygous and heterozygous for the transgene too) ▶️ allow study of dominant and recessive diseases