Gene cloning

Question 1

What is vector DNA?

Answer 1

DNA molecule that functions as a carrier to bring DNA into host cell and start replication

Question 2

What are artificial chromosomes?

Answer 2

Chromosomes made from genetic engineering

Question 3

What is in artificial chromosomes?

Answer 3

Centromere and telomere

Question 4

What is a DNA library?

Answer 4

Collection of DNA fragments stored in a population of microorganisms

Question 5

What does F factor stand for?

Answer 5

Fertility factor

Question 6

Where is F factor used?

Answer 6

DNA transduction

Question 7

What is the multiple cloning site?

Answer 7

Short sequence of DNA with restriction sites

Question 8

What are YACs?

Answer 8

Yeast artificial chromosomes which are 200-3000kb in size

Question 9

What are BACs?

Answer 9

Bacterial artificial chromosomes which are 100-300kb in size

Question 10

Where are YACs used?

Answer 10

Yeast cells

Question 11

Role of BACs

Answer 11

Transforming and cloning in bacteria

Question 12

Size of bacteriophages

Answer 12

90kb

Question 13

What are cosmids?

Answer 13

Plasmids containing DNA sequences from bacteriophage I

Question 14

Size of cosmids

Answer 14

45kb

Question 15

What are fosmids?

Answer 15

Like cosmids but based on bacteria plasmid containing F factor

Question 16

Size of bacteriophage I

Answer 16

9-23kb

Question 17

What are plasmids?

Answer 17

Small circular dsDNA

Question 18

What is an ORI?

Answer 18

Origin of replication

Question 19

Role of plasmids

Answer 19

Replicate independently of chromosomes

Question 20

What is phenotypic screening?

Answer 20

Finding the protein encoded by the gene that changes the colour of a colony

Question 21

What are bacteriophages?

Answer 21

Viruses that infect bacteria

Question 22

What type of molecules are bacteriophages?

Answer 22

ds molecules with ss ends

Question 23

Role of bacteriophages

Answer 23

Clone larger segments of DNA

Question 24

What is complementary DNA?

Answer 24

DNA synthesized from an mRNA

Question 25

What synthesizes mRNA into cDNA?

Answer 25

Reverse transcriptase and DNA polymerase

Question 26

How is DNA cloned?

Answer 26

Insertion of a foreign DNA fragment into a vector capable of replicating

Question 27

Purpose of replicating DNA

Answer 27

Isolating DNA DNA libraries Perform molecular studies

Question 28

What are the 4 methods for DNA cloning?

Answer 28

Restriction enzyme digestion and ligation T-A cloning TOPO attached unidirectional cloning Recombination based cloning

Question 29

How do DNA molecules get inside bacteria cells?

Answer 29

They are heat-shocked

Question 30

What is electroporating?

Answer 30

Using electric shocks to allow DNA to enter host cells

Question 31

What is conjugation?

Answer 31

Natural transmission from donor to recipient

Question 32

What is transfection of DNA?

Answer 32

Packing DNA into phage cells which then infect bacteria and so DNA is expressed

Question 33

What is a selectable marker?

Answer 33

Gene introduced into a cell that confers a trait for artificial selection

Question 34

Once DNA is cloned, what must be identified?

Answer 34

Transformed cells carrying the DNA

Question 35

What is the Gateway entry clone?

Answer 35

Silent clone used to shuffle DNA into expression vectors

Question 36

Purpose of Gateway entry clone

Answer 36

Allows the study of genes of interest