Chapter 11: Genetic Engineering and Biotechnology

Question 1

Restriction enzymes

Answer 1

Cut DNA at specific palindromic sequences

Question 2

Modification enzymes

Answer 2

Protect a cell’s DNA from its own restriction enzymes

Question 3

Hybridization

Answer 3

Single strands of DNA or RNA by matching of base pairs

Question 4

Southern blot

Answer 4

DNA is target, DNA or RNA are probes

Question 5

Northern blot

Answer 5

RNA is target, DNA or RNA are probes

Question 6

Western blot

Answer 6

Proteins are target

Question 7

FISH

Answer 7

fluorescence in situ hybridization

attach fluorescent probe to short single stranded DNA/RNA (oligonucleotides) to target specific DNA sequences

Question 8

PCR

Answer 8

Thermophilic DNA polymerase (like tag) amplifies DNA in cycles

Question 9

RT-PCR

Answer 9

Reverse transcription PCR
Make DNA from RNA templates
Start by converting RNA into cDNA

Question 10

qPCR

Answer 10

Quantitative PCR

Uses fluorescent probes to mintor amplification

Question 11

Steps of cloning

Answer 11

Cut foreign DNA with restriction enzyme
Cut vector (usually virus or plasmid or any other small element) with the same restriction enzyme
Use DNA ligase to bind foreign DNA with vectors
Introduce recombinant plasmid into host cell
Screen to ensure clone was introduced correctly

Question 12

Screening methods to ensure proper cloning

Answer 12

Antibiotic resistance markers
Southern blots
Antibodies that react with target protein

Question 13

Expression vector

Answer 13

Contains regulatory sequences that allow for manipulation of gene expression

Question 14

Reporter genes

Answer 14

Encode proteins that are easy to detect and assay

GFP