Genetic Engineering/Biotechnology (BE #6)

Question 1

What is recombinant DNA?

Answer 1

DNA that contains information from 2 different species of organisms.

Question 2

What are the 2 major goals of recombinant DNA technology involving the use of bacteria?

Answer 2

1. gene amplification - make lots of copies of gene of interest.
2. protein synthesis - get the bacteria to synthesize the protein coded for on the gene of interest.

Question 3

What is big difference between eukaryotic & prokaryotic chromosomes?

Answer 3

Eukaryotic chromosomes have noncoding (introns) sequences, while prokaryotic chromosomes have only coding (entrains) regions.

Question 4

Why can’t eukaryotic DNA be put directly into prokaryotic chromosomes?

Answer 4

Prokarytic chromosomes are unable to read the noncoding sequences that are present in eukaryotic DNA.

Question 5

cDNA is made in a process called __________, which is produced from mRNA.

Answer 5

reverse transcription

Question 6

What is the name of the enzyme that catalyzes reverse transcription?

Answer 6

reverse transcriptase

Question 7

cDNA can be inserted into a bacterial chromosome because it lacks the (coding or noncoding) sequences?

Answer 7

noncoding

Question 8

Order of cDNA & reverse transcription

Answer 8

DNA - pre mRNA - mRNA - mRNA/cDNA hybrid - denature to get SS cDNA - DS DNA

Question 9

What enzymes are synthesized by bacterial cells to cut apart & destroy foreign bacteriophage (viral) DNA molecules?

Answer 9

Restriction enzymes

These are used for the purpose of recombining genes.

Question 10

What are the resulting staggered cuts on the DNA called:

Answer 10

sticky ends

Question 11

T or F

In order to insert a eukaryotic gene into a prokaryotic plasmid, the same restriction enzyme must be used on both.

Answer 11

True

Question 12

What is a common method of DNA amplification?

Answer 12

Polymerase Chain Reaction (PCR)

Question 13

What does heat do to a double stranded nucleic acid molecule like DNA? What is this process called?

Answer 13

Heat breaks the hydrogen bonds between the 2 strands.

It’s called denaturing.

Question 14

Briefly explain the 3 steps of PCR.

Answer 14

1. Denaturing - heat DNA so strands split
2. Annealing - cool down & add primers; primers bind to single strands of DNA
3. Extension - warm up; add free nucleotides and DNA polymerase to finish replication. Original strands act as template.

Question 15

Is PCR done in vitro or in vivo?

Answer 15

in vitro (in a test tube)

Question 16

Give 2 examples of successful introduction & expression of a human gene by a bacterium.

Answer 16

1. vaccines
2. human growth hormone
3. insulin