In Vivo And In Vitro Cloning

Question 1

Isolating genes from human DNA

Answer 1

Use restriction endonuclease

To cut DNA at a specific base sequence

Question 2

Reverse transcriptase

Answer 2

Makes cDNA from RNA

Question 3

Restriction endonucleases

Answer 3

Enzymes that cut DNA into smaller fragments

Question 4

Recombinant DNA

Answer 4

Contains genes/sections of DNA

DNA from two species/2 types of organisms

Question 5

Sticky ends

Answer 5

Joins two pieces of DNA by complementary base pairing/binding

Question 6

Making recombinant DNA

E.g. making modified plasmids

Answer 6

Cut desired gene from another organism / mRNA from cells / organism.
Using restriction endonuclease / reverse transcriptase to get DNA (if it is mRNA)
Use restriction endonuclease to produce complementary sticky ends
Using the same enzyme to cut the plasmid/gene
Mix together and use DNA ligase to join wanted gene to plasmid/ other gene (at sticky ends)
(If a plasmid) - return to cells and place in solution of Ca2+ which makes the bacteria cell walls and membranes more permeable, allowing plasmid to be taken up.

Question 7

Identification

Answer 7

Plasmids may contain gene that codes for fluorescence protein
Let’s them glow under UV light
Allows quick identification of transformed bacteria under UV light
Plasmids may contain antibiotic resistance genes
They act as a marker gene
This allows detection of cells containing plasmid/DNA

Question 8

Vector

Answer 8

E.g. Plasmid
A carrier of DNA / gene
Into a cell / other organism / host

Question 9

DNA probe

Answer 9

Single stranded DNA
Base sequence complementary to DNA / gene that is being identified
Radioactively / fluorescent labelled so it can be detected