Ch 20

Question 1

recombinant DNA

Answer 1

DNA molecules formed when segments from different sources are combined in vitro

Question 2

in vitro

Answer 2

in a test tube

Question 3

biotechnology

Answer 3

manipulation of organisms or their components to make useful products

Question 4

genetic engineering

Answer 4

direct manipulation of genes

Question 5

DNA Cloning

Answer 5

production of copies of defined DNA segments

Question 6

plasmid

Answer 6

small circular SNA molecules that replicate separately form the bacterial chromosome

Question 7

Steps of DNA Cloning

Answer 7

1. gene inserted into plasmid
2. plasmid put into bacterial cell
3. host cell reproduces through cell division to form a clone of cells

Question 8

clone

Answer 8

population of genetically identical cells

Question 9

gene cloning

Answer 9

production of copies of a single gene

Question 10

amplify

Answer 10

to make many copies of a gene

Question 11

What do restriction enzymes do?

Answer 11

cut DNA molecules

Question 12

restriction/recognition sites

Answer 12

specific sequence where a restriction enzyme can cut DNA into restriction fragments

Question 13

What is special about a restriction site?

Answer 13

the sequence is a palindrome

Question 14

sticky end

Answer 14

single-stranded end of fragment where it can attach to others

Question 15

DNA ligase

Answer 15

enzyme that seals sticky ends together

Question 16

isoschizomer

Answer 16

different restriction enzymes that produce the same cut

Question 17

cloning vector

Answer 17

DNA molecule that can carry foreign DNA into a host cell and replicate

Question 18

Set up for cloning genes in a bacterial plasmid

Answer 18

1. Obtain gene of interest and a plasmid with a resistance gene
2. Cut with the same restriction enzyme. Plasmid will be cut in the lac z gene.
3. Mix plasmids and fragments; some will join.
4. DNA mixture added to bacteria

Question 19

Steps to identify the correct bacterial plasmid

Answer 19

Place bacteria in a medium with ampicillin and x-gal.
Only bacteria with the plasmid will reproduce because they are amp resistant.
Only bacteria containing a recombinant plasmid/insert in lac z will turn white because they can’t hydrolyze x-gal.

Question 20

cDNA

Answer 20

complementary, double stranded DNA formed form mRNA

Question 21

Enzyme for cDNA synthesis

Answer 21

reverse transcriptase

Question 22

cDNA shows only…

Answer 22

expressed genes

Question 23

Purpose of Nucleic Acid Hybridization

Answer 23

detect specific sequence by its ability to base pair with a complementary DNA

Question 24

Steps of Nucleic Acid Hybridization

Answer 24

1. DNA transferred to a nylon membrane.
2. Incubated in a solution of radioactive probe molecules that will base pair with complementary DNA.
3. Excess DNA is rinsed off.
4. Covered with film that exposes radioactivity

Question 25

What does PCR stand for?

Answer 25

polymerase chain reaction

Question 26

purpose of PCR

Answer 26

amplify a sequence through in vitro replication

Question 27

Steps of PCR

Answer 27

1. Denature
2. Anneal
3. Extend

Question 28

enzyme of PCR

Answer 28

Taq DNA polymerase to extend and to last through denaturing

Question 29

Gel electrophoresis

Answer 29

gel separates nucleic acids or proteins by size.

Question 30

In gel electrophoresis, ________ moves __________.

Answer 30

Shorter moves faster

Question 31

RFLP

Answer 31

loss of a restriction site = change in band pattern

Question 32

Southern blotting

Answer 32

detects only bands with part of a specific gene

Question 33

Steps of southern blotting

Answer 33

1. gel electrophoresis
2. transfer/blotting to membrane
3. hybridization

Question 34

Northern blotting

Answer 34

same as Southern but mRNA instead of DNA

Question 35

Steps of Dideoxy chain termination

Answer 35

1. Denature strands and anneal primers
2. strands synthesized until a dideoxyribonucleotide is added
3. Read the tagged nucleotides