Biotechnology Tools and Techniques

Question 1

Biotechnology tools and tecniqs
Manipulating DNA:
Recombinant DNA?
Biotechnology tools and techniques?(4)

Answer 1

Recombinant DNA- a fragment of DNA composed of sequences from at least two different sources

Restriction enzyme, methylases, ligase, gel electrophoresis

Question 2

Biotechnology tools and tecniqs
Restriction enzyme?
What do they do?
How do they do the cutting?
Function?

Answer 2

• molecular scissors
• recognize specific base-pair sequences in DNA and then cuts the double-stranded DNA at those sites
• binds to recognition site
• hydrolysis reaction to disrupt phosphodiester bond
• H bond between bases disrupted
• results in two fragments of DNA to work with
• rudimentary immune system in bacteria
• cleaves virus DNA into fragment
• host DNA is methylated, RE knows not to cleave

Question 3

Biotechnology tools and tecniqs
Restriction enzymes continued:
Recognition site? Characteristics?
Sticky ends?
Blunt ends?
Which is more useful for biotechnology? Why?

Answer 3

-The sequence recognized by the enzyme 
Characteristics: 
-specific to each Restriction enzyme
-4-8 base pairs in length, allows for low frequency of cuts
-8 base pairs RS = to long 
-6 base pairs RS = ideal
-palindromic 

• cleavage produces an overhang
• depending on where the restriction enzyme cuts, can be 5’ or 3’ overhang
• no overhang
• sticky ends because if two fragments cut with same restriction enzyme they will have complementary ends and then fragments can be joined together easily

Question 4

Biotechnology tools and tecniqs
Methylases?
Function?
As biotechnological tool?

Answer 4

• methylases are enzymes
• add methyl group to the recognition site
• prevent restriction enzymes from cleaving the DNA
• protects host from own restriction enzymes
• allow protection of fragments/specific sequences

Question 5

Biotechnology tools and tecniqs
Ligases joins?
Used to join fragments that have?

Answer 5

• sugar and phosphate back bones of DNA fragments

- fragments that have complementary ends and phosphodiester bonds

Question 6

Biotechnology tools and techniques

What is biotechnology?

Answer 6

Applied biology- genetics, molecular biology, microbiology, biochemistry

Uses living organisms and their components to creat “bio-products”- industry, agriculture, medicine and involves the manipulation of DNA