Exam 3 Part 3

Question 1

genetic engineering

Answer 1

human manipulation of an organism’s DNA

Question 2

genetic engineering methods

Answer 2

recombinant DNA tech

Question 3

recombinant DNA technology

Answer 3

transplating a gene from one organism to another
rDNA is created using molecular cloning
DNA of interest is inserted into a cloning vector

Question 4

what would the final clone be used for?

Answer 4

used the gene product
find what the gene product does
diagnosis of a disease
gene therapy
mutation
sequence

Question 5

strategy of genetic engineering

Answer 5

1. isolate DNA of interest from an organism
2. cut the DNA with a restriction enzyme
3. cut the cloning vector with the same restriction enzyme
4. ligate piece into a cling vector to make a recombinant DNA molecule
5. transform the recombinant DNA into a host (like e. coli) the host cell will make identical copies called clones

Question 6

restriction enzymes

Answer 6

produce sticky ends

naturally occuring

Question 7

genomic library preparation

Answer 7

DNA is extracted from the organism of interest
cut DNA into small pieces using restriction enzymes
ligate pieces into cloning vector
form library

Question 8

genomic library

Answer 8

collection of clones/plasmids in which each plasmid contains a different piece of DNA

Question 9

why have genomic libraries?

Answer 9

screen libaray to look for a specific sequence
diagnostics
squence by shot gun screens

Question 10

screening a genomic library

Answer 10

goal: find a DNA sequence in a pool of DNA
use a DNA probe
caveat

Question 11

DNA probe

Answer 11

detectable tag on a primer

Question 12

cDNA library prepartion

Answer 12

RNA is extracted from an organism of interest
RNA is reverse transcribed into cDNA
Each cDNA is cloned into vector
Each library clone has a different cDNA molecule

Question 13

why have cDNA libraries?

Answer 13

Detect sequence of interest
tell difference bt exons and introns
splice varients

Question 14

splice variants

Answer 14

can tell where exons/introns are-coding regions

Question 15

uses for PCR

Answer 15

amplify a DNA segment of interest
forensics
organ matching
sequence an entire genome
quantify a transcript
make mutations

Question 16

Real-time PCT

Answer 16

quatitative PCR
measure the increase in the amount of PCR product
measure the abundanence of a transcript in a sample
measure cDNA as it is amplified

Question 17

PCR to inroduct mutations

Answer 17

study mutation
identify a disease
identify the important amino acid in a polypeptide
to understand the role of a specific amino acid in a polypeptide

Question 18

types of mutagenesis

Answer 18

site-directed mutagenesis

random mutatgenesis

Question 19

site directed mutagenesis

Answer 19

mutate one specific nucleotide

include primer with single nt change

Question 20

random mutagensis

Answer 20

create random mutatiosn ian DNA of interest
add Mn2+ instead of Mg2+
causes polymerase to have reduced fidelity

Question 21

Southern blotting

Answer 21

detect a specific DNA sequence in a sample
used to compare homologous genes
analyze intron organization
paternity tests
diagnostics
methylated sequences

Question 22

Southern blotting procedure

Answer 22

1. isolate DNA from organism of interest
2. cut into small pieces using restriction enzymes
3. sparate DNA fragments using gel electrophoresis
4. transfer DNA from gel to a membrane/blot
5. incubate blot with a labelled probe that is complementary to the DNA of interest
6. visualize the probe–band will be DNA of interest