13 - recombinant DNA

Question 1

recombinant DNA

Answer 1

Recombine DNA from two distinct sources. (DNA fragment of interest + plasmid vector)

Question 2

Conclusion of griffiths experiment

Answer 2

A chemical substance from one cell is genetically transforming another cell

Question 3

restriction enzyme

Answer 3

cut DNA into small fragments, endonucleases, recognise restriction sites, palindromic, make sticky ends and blunt end cuts (staggered or complementary)

Question 4

Vector

Answer 4

a DNA molecule that replicates within a living cell, eg plasmid or virus. Contain genetic signals for replication, inserting foreign DNA and identifying cells that contain recombinant DNA

Question 5

Types of vectors

Answer 5

• Plasmid
• Phage lambda (bacteriophage)
• Cosmid
• Bacterial artificial chromosome

Question 6

Plasmid

Answer 6

Circular dna molecule that replicates seperate from chromosome DNA. Occur naturally in bacteria and are replicated before every cell division

Question 7

What is a plasmid polylinker

Answer 7

contains recognition sequence for restriction enzymes

Question 8

DNA ligase

Answer 8

catalyses phosphodiester bond formation, requires ATP

Question 9

How to get recombinant DNA into cell?

Answer 9

Transformation. The uptake of exogenous DNA through the cell membrane. Bacterial cells that take up plasmid can be selected

Question 10

Transforming principle experiment

Answer 10

• Virulent S strain bacteria = mouse dies
• Non virulent R strain bacteria = mouse lives
• heating kills virulent s strain bacteria = mouse lives
• mix dead s strain bacteria and living non virulent = mouse dies

Question 11

Library

Answer 11

Collection of N vectors with different inserted fragments of DNA

Question 12

3 types of gene libraries

Answer 12

• DNA library
• Genomic library
• cDNA library

Question 13

Genomic library

Answer 13

Clones that collectively represent all DNA sequences of the genome

Question 14

DNA Library

Answer 14

Collection of clones containing all the DNA fragments

Question 15

cDNA library

Answer 15

Clones representing all mRNA in a cell (mRNA cloned as cDNA into vector)

Question 16

Screening DNA libraries

Answer 16

DNA libraries can be screened by hybridisation to a DNA probe

Question 17

Uses of recombinant DNA tech

Answer 17

Pharamaceuticals, medical testing, research, GM food, transgenic animals

Question 18

Glofish

Answer 18

Insertion (recombinant plasmid injected into cytoplasm of 1 or 2 cell stage embryos)
screening (protein expression began) resulting in glowing fish

Question 19

Vector for glofish

Answer 19

plasmid pEGFP-1

Question 20

Restriction enzymes for GloFish

Answer 20

EcoR1

Question 21

Cloning alternative

Answer 21

Use PCR to make copies of DNA fragment instead of E.coli

Question 22

Genome engineering

Answer 22

Methods that allow changes to be introduced into DNA of many organisms by inducing a targeted double strand break into the genome