Genetic Technologies

Question 1

Copy Number

Answer 1

of copies of a plasmid allowed in a cell

Question 2

Conjugation Tube

Answer 2

Tube between two bacteria to share plasmids

Question 3

Vectors

Answer 3

any particle used as a vehicle to artificially carry a foreign nucleic sequence – usually DNA – into another cell

Question 4

Process of Vector modification

Answer 4

1) Endonucleases open plasmid
2) New DNA added
3) Plasmid loop closed

Question 5

Restriction Enzymes

Answer 5

Made by bacteria to degrade foreign DNA

Question 6

Restriction Sites

Answer 6

Specific places where restriction enzymes cut (usually 4-6 base pairs)

Question 7

Prevention of self-degradation of double-stranded DNA

Answer 7

Methylation of host DNA

Question 8

Type II Restriction Enzymes

Answer 8

Most useful for genetic engineering

Question 9

Palindrome

Answer 9

DNA sequence that’s complement is the same read 5’ -> 3’

Question 10

What type of DNA is utilized specifically for type II restriction enzyme sequences?

Answer 10

Palindromic DNA

Question 11

Sticky (Staggered) Ends

Answer 11

Cut DNA that looks “staggered” (SAME ONES can be brought together and religated)

Question 12

Blunt Ends

Answer 12

Ends that line up on their base pairs (Any blunt ends can pair up thru DNA Ligase)

Question 13

Recombinant DNA

Answer 13

DNA from two organisms brought together

Question 14

Reqs of a Plasmid Vector Molecule

Answer 14

1) Has an ori
2) Has restriction sites for enzyme used
3) Has some way to select for the vector in the cell
4) Distinguish between vector and vector + cell

Question 15

Reporter Gene

Answer 15

Gene that “reports” whether or not a plasmid has been changed, knocked out upon insertion of DNA of interest

Question 16

Polylinker

Answer 16

Collection of UNIQUE restriction sites

Question 17

Transduction

Answer 17

Phage carries DNA into bacterial cell

Question 18

Transformation

Answer 18

Make a cell transiently permeable to DNA in its surroundings, s/t the cell intakes the DNA

Question 19

Cell Clones

Answer 19

Cells that carry copies of the DNA of interest (Recombinant DNA)

Question 20

Purpose of Cloning

Answer 20

AMPLIFICATION

Question 21

Genomic Library

Answer 21

Entirety of an organism’s genome chopped up into pieces by restriction enzymes

Question 22

Preparation of genomic library

Answer 22

1) Chop genomic DNA w/ restriction enzymes
2) Attach fragments to vectors
3) Introduce E.Coli to amplify amount of vectors

Question 23

cDNA Library Preparation

Answer 23

1) Harvest appropriate tissue, isolate total processed mRNA representing genes expressed by a particular cell type
2) Use Reverse Transcriptase to make ss-DNA (Mutation rate: 1/2000)
3) Use DNA Poly to make complementary strand (prevents mutation rate, called cDNA)
4) cDNA inserted into vector, transform in host cells
5) Gene transcribed/translated (cDNA has no SD sequences or promoter regions, vector molecule has that)

Question 24

cDNA library vs. Genomic Library

Answer 24

Genomic library = ENTIRE GENOME
cDNA library = ONLY PROTEINS BEING MADE AT THAT TIME

Question 25

Polymerase Chain Reaction (PCR)

Answer 25

Limited by knowledge about surrounding bases 1) Custom primer anneals to target sequences 2) Heat-stable DNA poly synthesizes rest of DNA 3) Rinse and Repeat! Creates millions of copies