Gene technology

Question 1

What is in vivo?

Answer 1

Made inside living organism

Question 2

What is in vitro?

Answer 2

MAde outside living organism

Question 3

How is reverse transcriptase used?

Answer 3

mRNA with gene is template

First isolate mRNA with gene

mRNA, free DNA nucleotides and reverse transcriptase is mixed

Nucleotides would align next to their complementary base on mRNA template

Then reverse transcriptase joins the DNA nucleotides together producing fragment of DNA

This fragment is called complementary DNA(cDNA)

Double strand produced when cDNA and DNA polymerase used together

Question 4

How is restriction endonuclease used?

Answer 4

It cuts DNA molecule at specific base sequence

It breaks sugar phosphate backbone of double stranded DNA

This results in either of sticky ends or bluent ends

Question 5

What is sticky ends?

Answer 5

Enable the DNA to be joined or spliced onto a different piece of DNA effectively to staggered cut

Form hydrogen bonds with complementary bases on other sticky end

Preferred

Question 6

What is blunt ends?

Answer 6

Can be used but there is more chance of joining non specific fragments

Question 7

How is DNA ligase used?

Answer 7

Joins foreign DNA into plasmid DNA

It rejoins the break between the nucleotides of each strand by reforming sugar phosphate backbone

DNA produced can now be replicated and multiplyied