Biotechnology

Question 1

Define recombinant DNA technology

Answer 1

• Formation of new combinations of genes
• By isolating genes from one organism
• And introducing them into similar or unrelated organism

Question 2

Tools used in recombinant DNA technology

Answer 2

• R.E
• Cloning vectors
• Hosts
• Ligase

Question 3

What is R.E a.k.a Endonucleases?
Function of R.E?

Answer 3

• Degradative enzyme in many bacteria
• Protect the bacterial cell cutting up foreign DNA from other organism or phages

Question 4

Why restriction endonucleases will not cleave the bacterial cells’ own DNA?

Answer 4

• Protection via METHYLATION addition of methyl groups (-CH3) to Adenines and Cytosines within the sequences recognized by enzymes

Question 5

How RE cut DNA?

Answer 5

• RE is very specific
• RE recognizes sequence 4-8 nucleotides.
• cleaves both strand at a specific point within restriction site
• Different RE = Different restriction site
• Rxn site: palindrome
• sequence symmetrical
• sequence of nucleotides is the same on both strands when read in the 5’ > 3’ direction

Question 6

Examples of restriction enzymes

Answer 6

EcoRI and SmaI

Question 7

EcoRI produces ___ ends

Answer 7

Sticky ends

Question 8

SmaI produces ____ ends

Answer 8

Blunt ends

Question 9

What is sticky end?

Answer 9

• Single stranded end of DNA

Question 10

Importance of sticky end

Answer 10

• can pairs/ complements with the other sticky end which has been cut with the same RE
• Enable to bind fragments from different sources (DNA fragment from (TAK SIAP LAGI)