Recombinant Proteins 2

Question 1

Where’s is the DNA sequence of interest propagated from?

Answer 1

In a cloning vector

Bacterial plasmid DNA
bacterial viruses
Yeast plasmids
Animal plasmids

Question 2

What are the barriers and solutions of making recombinant proteins?

Answer 2

Bacteria cannot process introns
And bacterial RNA polymerase cannot recognise mammalian promoter sequences

Solutions: remove intron sequences or use cDNA (copy of processed mRNA)
Insert into a vector behind a bacterial promoter

Question 3

How to make cDNA from RNA templates?

Answer 3

Reverse transcriptase + poly dT primer

Degrade RNA

DNA polymerase + random primer

Question 4

What is PCR?

Answer 4

Polymerase chain reaction

Amplification of a specific DNA sequence

Question 5

What is a primer?

Answer 5

A short DNA sequence usually 15-30 base pairs long

Question 6

Can restriction enzymes sites be used to insert into PCR primers?

Answer 6

Yes

Question 7

In the Sanger sequencing why is Dideoxy NTPs inserted?

Answer 7

To cause chain termination

Question 8

What are the advantages of inducible expression vectors?

Answer 8

Controlled expression
Maximise yield and solubility
Minimise toxicity to host cell
Inert without addition of inducer

Question 9

What can recombinant proteins be produced from?

Answer 9

Bacterial cells 
Yeast cells 
Cultured mammalian cells 
Transgenic plants and animals
In vitro systems