Biotechnology

Question 1

Define Biotechnology

Answer 1

The industrial application of microorganisms, cells, or cell components to make a useful product.

Question 2

Define rDNA

Answer 2

A DNA molecule produced by combinign two or more sources of DNA.

Question 3

What are the steps of a typical genetic modification procedure?

Answer 3

1. Obtain a Vector
2. Obtain a GOI
3. Insert GOI into Vector; Insert rDNA into baterium
4. Tranform Bacteria expressing GOI
5. Clone
6. Collect GOI in large numbers OR
7. Collect GOI products and use

Question 4

What are Restrictive Enzymes?

Answer 4

• A special class of DNA-cutting enzymes that exist in many bacteria which recognizes and cuts only one particular sequence of nucleotide bases in DNA, and it cuts this sequence in the same way each time.
  
  • Blunt ends
  • Sticky Ends
• Can be used to cut GOI and cut receipient DNA molecule for insertion of GOI from another organism.

Question 5

Define Vector

Answer 5

A vector is a way to hold a GOI or DNA sequence of interest from one organism and it is used to insert the GOI into ANOTHER organism.

Question 6

What are the Requirments to be a Vector?

Answer 6

1. Must be self-replicating
2. Small size
3. Must contain markers to correctly identify organism that acquired the vector
4. Common vectors include plasmids & viruses

Question 7

What is a Polymerase Chain Reaction?

Answer 7

A way to make multiple copies of DNA sequence very quickly.

Question 8

What are different techniques used to insert foreign DNA into cells?

Answer 8

• Cell to cell contact
  
  • Ex. conjugation
• Transformation
  
  • ex. Grith ex
• Electroporation
  
  • the use of electrical current to create microscopic pores so foreign DNA can enter.
  • If cell contains cell wall then convert to protoplast first by using enzymes.
• Microinjections
  
  • Ex. Directly into plant or animal cell cytoplasm

Question 9

What are different techniques used to select clones?

Answer 9

• Blue-White Screening
  
  • Plasmid inserted with markers ampR and LacZ
  • Placed into X-gal medium, which contains ampicilin and B-galactosidase
  • If amp-R gene transffered, bateria will survive
  • If bacteria picked up LacZ they will hydrolize X-gal and turn blue.
  • Foreign Bacteria with GOI will turn White
• Colony hybirdization
  
  • DNA Probes
    
    • attach to target gene if present and serve as markers.

Question 10

How is human insulin produced using biotechnology?

Answer 10

1. Synthetic genes produced for the two short strands of polypeptide chains that make up insulin
2. Insert into plasmid vector and link to LacZ
3. Two different e.coli bacteria are used for each insulin strand
4. Recover insulin strands from bateria and seperate from LacZ
5. Join the two strans of insulin to produce Human insulin