21 - Recombinant DNA technology

Question 1

What is a GM organism?

Answer 1

An organism that has had its DNA altered through recombinant DNA technology.

Question 2

Describe the creation of DNA fragments using restriction endonuclease.

Answer 2

Cut at recognition sites near the desired gene.

Question 3

Describe the creation of DNA fragments using reverse transcriptase.

Answer 3

mRNA of desired gene converted to complementray DNA (cDNA) using reverse transcriptase

Double stranded DNA synthesised using taq DNA polymerase

Question 4

Describe the creation of DNA fragments using a gene machine.

Answer 4

gene sequence determined using primary protein structure

Question 5

Describe the process of in vivo DNA amplification

Answer 5

1. DNA fragmented
2. Promoter and terminator region added
3. Gene added to plamid using restriction endonuclease to create sticky ends in plasmid which join as they are complementary. Joined by DNA ligase
4. Transformation (plasmid into cell)

Question 6

Describe the use of marker genes.

Answer 6

• Plasmids contain two marker genes
• One shows antibiotic resistance so the bacteria can be identified as having taken up a plasmid
• Second is a phenotypic gene that is split by the target gene being taken up making it non-functional
• This can be a fluoresence gene, a different antibiotic resistance gene or an enzyme action gene

Question 7

what does PCR stand for?

Answer 7

Polymerase chain reaction

Question 8

What does PCR require?

Answer 8

10,000 base pairs
nucleotides
taq DNA polymerase
complementary primers
thermocycler

Question 8

What does PCR require?

Answer 8

10,000 base pairs
nucleotides
taq DNA polymerase