Strain Development and Recombinant DNA Technology

Question 1

Basic Steps in Strain Development for industrial fermentations

including mutagenesis and selection of desireable mutants

Answer 1

Goal: Increase the yield of a desired product
General Idea: Change Genotype and Select strain with increase yield

Question 2

Changing Genotype

2 methods

Answer 2

• Use mutagenic agents to bring genomic changes.
  Common Mutagens: Short wave UV light, HNO2, NH2OH
• Use recombinant DNA techniques

Nitrous Acid, Hydroxylamine

Question 3

Mutagenesis

Expected use/performance

Answer 3

Can’t be accurately predicted. Develop dose-response curve.

Question 4

Selection of Desirable Mutants

2 methods

Answer 4

Random Screening
Selective Isolation

Question 5

5 requirements involved in recombinant DNA technology

corresponding tools as well

Answer 5

Isolate Gene
Cutting Dna
Joining Dna
Introducing Dna into host cells
Selecting host cells expressing gene

Question 6

Polymeraze Chain Reaction

Purpose and Requirements

Answer 6

Purpose: Small amount of dna –> larger amount of dna
Requires: DNA amplification, DNA polymerase, 2 oligonucleotide primers

Question 7

PCR Procedure

Answer 7

1. Heat the double stranded DNA to separate strands
2. Cool reaction mixture so oligonucleotide anneal to sequence
3. Synthesize the DNA with DNA polymeraze
4. Repeat 1-3 until desired DNA quantity is obtained

Question 8

Transgenic Animals

Definition and development

Answer 8

Animals that have had a foreign gene instered into their genome
Microinject gene into fertilized eggs and implant egg in mature female

Question 9

CRISPR is a powerful ____ tool

Hint: 2 words

Answer 9

gene editing