biotech

Question 1

exons

Answer 1

coding sequence of proteins but usually dispersed within introns

Question 2

cloning

Answer 2

process of creating exact copy of biological unit

• defined sequence of dna

Question 3

gene

Answer 3

specefic sequence of dna that should be unchanged when copied

Question 4

reverse transcriptase

Answer 4

turn mrna back into dna
turns dna into pre-mrna then cellular factors cut out introns and splice exons togerher

Question 5

4 stages of biotech

Answer 5

1. ISOLATE gene from donor dna (chromkosome from b-pancreatic cell)
   2.INSERT gene into suitable cloning vector (bacterial plasmid)
   3.introduce VECTOR into an expression host (bacteria or virus transcribe into mrna to translate into human insulin)
   4.use recombinant expression host to scale up and purify, formulate, QC

Question 6

PCR

Answer 6

1.denaturation - reverse transcriptase to seperate dna strands
2.annealing - 2 strands cooled and joined together by primers
3. synthesis - dna polymerase works on both strands
4.now repeat for 2 dna strands and keep replicating 2-4-8-16 so on

Question 7

cloning vector

Answer 7

small piece of dna taken from a a virus, a plasmid or cell of higher orgainism - into which a foreign dna fragment can be inserted for cloning

Question 8

plasmid

Answer 8

small double stranded self replicating circular dna molecules, distinct from normal bacterial chromosone

Question 9

process of cloning

Answer 9

1. insert gene into cloning vector
2. transfect chimeric vector into prokaryotic cell (e-coli) the transformed
   3.screen transformed cells to ensure correct gene

Question 10

how to transform cell

Answer 10

1. make cell temporarily permeable (competent) so that it can be take up plasmid dna
2. the stop permeability so it doesnt leak back out

acheived witch heatshock (treated with CaCl heated to 42C then 0C) or electrotransformation (shocked with 200-2000V for a few minutes)

Question 11

expression system

Answer 11

determines what its used for

Therapeutic antobidy must have full biological functionality to be effective

expressiopn system must oerform all required post tranlational modifications so replicated protein resembles native protein.

Question 12

protein processing

Answer 12

additional modification after proteins have been translated to become fully functional (eg when proteins are glycosylated)