Recombinant DNA Technology

Question 1

substitution mutation

Answer 1

one base swapped for another

Question 2

insertion/ deletion mutations

Answer 2

one base removed or added

causes a frame-shift and completely alters the protein being synthesised

Question 3

conservative substitution mutation

Answer 3

base codes for aa with similar properties so can be tolerated

Question 4

non-conservative substitution mutation

Answer 4

base codes for different aa with different properties, not tolerated- diseases

Question 5

silent substitution mutation

Answer 5

new base codes for the same aa meaning no change

Question 6

gene cloning

Answer 6

produces copies of a particular DNA stretch

Question 7

gene cloning method

Answer 7

• desired gene isolated with restriction enzymes
• gel electrophoresis
• desired gene inserted into plasmid
• plasmid introduced to bacterium and allowed to grow therefore multiplying the gene

Question 8

restriction enzymes

Answer 8

cut double stranded DNA at specific DNA sequences (their recognition point)

Question 9

sticky ends

Answer 9

staggered cut of restriction enzymes

Question 10

palindromic

Answer 10

same backwards and forwards

Question 11

gel electrophoresis

Answer 11

separates DNA fragments based on their size

charge applied to gel , large fragments migrate slower

Question 12

DNA sequencing

Answer 12

used to determine base sequences of DNA

Question 13

Sanger sequencing

Answer 13

old method for DNA sequencing

synthesis of new DNA strand complementary to a single stranded template

Question 14

dNTPs

Answer 14

deoxynucleotides - DNA bases

Question 15

ddNTPs

Answer 15

modified nucleotides that terminate DNA strand elongation

3’ hydroxyl group is replaced with a H atom meaning it can’t form phosphodiester bridge

Question 16

Sanger sequencing procedure

Answer 16

1 - desired DNA strand and primer mixed
2 - mixture divided into 4 tubes containing , all 4 dNTPS, one of 4 ddNTPs , DNA polymerase
3 - chain synthesis occurs in each tube
4 - gel electrophoresis shows chain termination location
5 - bands read from bottom to top to deduce sequence

Question 17

next generation sequencing

Answer 17

can sequence an entire genome at once

very fast and cheap