Recombinant DNA Technology

Question 1

Who won the Nobel Prize for the first recombinant DNA experiment?

Answer 1

Paul Berg
(1980)

Question 2

What is Transformation?

Answer 2

Genetic alteration of a cell resulting from the direct uptake and incorporation of exogenous genetic material from its surroundings

Question 3

How many types of Restriction Enzyme are there?

Answer 3

3

Question 4

Which type of restriction enzymes are most often used in genetic engineering?

Answer 4

Type II

Question 5

What modification protects DNA sites from restriction endonuclease?

Answer 5

Methylation

Question 6

Who made the observation that phage grown in one bacterial host fail to grow in a different “restrictive” bacterial host?

Answer 6

Salvador Luria

Question 7

Who defined the molecular basis of restriction and modification?

Answer 7

Werner Arber
(1962)

Question 8

Who characterised the first restriction endonuclease?

Answer 8

Matt Meselson & Bob Yuan

Question 9

What DNA ligase is used in recombinant DNA?

Answer 9

T4 DNA Ligase

Question 10

What are the essential features of a plasmid vector?

Answer 10

1. Origin of Replication
2. Antibiotic Selection
3. Multiple Cloning Sites

Question 11

Who first indentified EcoRI?

Answer 11

Herbert Boyer

Question 12

How do Restriction Enzymes work?

Answer 12

Cut dsDNA at specific sequence creating blunt or sticky ends

Question 13

What was the first recombinant DNA experiment?

Answer 13

Inserting new genetic information into DNA of simian virus 40 to produce circular SV40 DNA

Question 14

What genes does circular SV40 DNA contain?

Answer 14

A molecule containing lambda phage genes and galactose operon of E. coli

Question 15

What was the first cloning experiment undertaken?

Answer 15

Cloning of frogs

Question 16

What was organised in 1975 due to fear of recombinant DNA technology?

Answer 16

A landmark conference in Asimolar California

Question 17

What are the THREE tools of molecular biology?

Answer 17

1. Transformation
2. Restriction enzymes & Ligase
3. Plasmids

Question 18

Is DNA parallel or anti-parallel?

Answer 18

Anti-parallel

Question 19

What are bacterial cells that can take up DNA known as?

Answer 19

Competent cells

Question 20

What are some bacteria that are naturally competent?

Answer 20

Pneumococcus & Neisseria

Question 21

How can E. coli become competent?

Answer 21

CaCl2 treatment and electroportation

Question 22

How many enzymes are commercially available?

Answer 22

More than 240

Question 23

How are restriction enzymes named?

Answer 23

Named for the organism they were discovered

Question 24

What are pallindromic sequences?

Answer 24

DNA sequence that is the same forward and backwards but on opposite strands

Question 25

What type of DNA fragment ends are more commonly used in genetic engineering and why?

Answer 25

Sticky ends because of their complimentary tails

Question 26

What is the size range of plasmids and do they carry essential genes?

Answer 26

1kb to 1.5Mb
They don’t carry essential genes, they carry virulence factors like antibiotic resistance genes

Question 27

What was the first plasmid vector?

Answer 27

pSC101
(SC = Stanley Cohen)

Question 28

What was the first biotechnology company?

Answer 28

Genentech

Founded by Herbert Boyer

Question 29

What was the first recombinant DNA experiment using restriction enzymes?

Answer 29

Newly constructed plasmid inserted into E. coli by transformation

Question 30

In what plasmid were synthetic genes for human insulin cloned?

Answer 30

In plasmid pBR322

Question 31

What are genomics?

Answer 31

Identify gene variants in populations

Question 32

What are the TWO enzymes needed for gene cloning?

Answer 32

1. Restriction enzymes
2. DNA ligase