Recombinant DNA Technology

Question 1

What is the significance of the enzyme Taq polymerase? What technique is it used for?

Answer 1

Acts as the DNA polymerase in PCR because it can handle the high temps in the process

Question 2

define cDNA

Answer 2

the DNA strand made by reverse transcriptase

Question 3

How are synthetic nucleic acids used?

Answer 3

used to make a DNA or RNA molecule using the precursors and enzymes. Used for determining a genome sequence, synthesizing a gene based on a protein sequence, synthesizing probes to isolate a specific sequence, and synthesizing mRNA that codes for a viral protein.

Question 4

Define RFLPs

Answer 4

restriction fragment length polymorphism. a collection of DNA fragments that are produced when treating DNA with restriction enzymes

Question 5

Define GMOs

Answer 5

an organism who’s genetics have been modified to improve it.

Question 6

Define exons

Answer 6

the protein-encoding parts of DNA

Question 7

How are gene libraries created? How is this process different for eukaryotic gene libraries?

Answer 7

the genome of an organism is split up gene by gene and put into vectors. Eukaryotic gene libraries are created from cDNA from mRNA to make sure we only get exons

Question 8

Define Southern blotting

Answer 8

can be used to probe a particular sequence of DNA. DNA is cut using restriction enzymes and then placed in an electric field in gel and fragments separate by length. Fragments are transferred to a nitrocellulose and tested for hybridization with a tagged probe.

Question 9

Define gene library

Answer 9

a collection of vectors containing single genes

Question 10

Briefly describe how to sequence DNA. [dideoxy (Sanger) sequencing especially]

Answer 10

copies of the DNA are made w/ four different solutions each containing a small amount of each ddATP (ATGC versions). Those ddATP bind to their corresponding base. This allows us to see where each base is in the sequence.

Question 11

What is a palindrome?

Answer 11

read the same back as forward

Question 12

Describe some applications of recombinant DNA technology.

Answer 12

medicine, agriculture, police work, legal stuff

Question 13

define introns

Answer 13

the non protein-encoding parts of DNA

Question 14

What are some typical products produced by recombinant DNA procedures?

Answer 14

foods, vaccines, antibiotics, hormones, proteins

Question 15

Describe four medical products that are genetically engineered.

Answer 15

insulin, tPA, aplha interferon, and hep b vaccine.

Question 16

Define PCR

Answer 16

a tool used to make copies of DNA, amplify the DNA

Question 17

What is the role of the vector in recombination?

Answer 17

a DNA molecule (often plasmid or virus) that is used as a vehicle to carry a particular DNA segment into a host cell as part of a cloning or recombinant DNA technique

Question 18

Define DNA fingerprinting

Answer 18

Originally was mattching patterns of bands that corresponded with RFLPs. Used for crime things and paternity.

Question 19

Describe 5 ways of introducing DNA into cells.

Answer 19

transformation- naked DNA
electroporation- holes in cell wall
protoplast fusion- cell wall removed by lysozyme and cell membranes merge
gene gun- DNA coated bullets shot into plant cell walls
microinjection- microscopic glass pipets inject DNA into cell

Question 20

What is reverse transcriptase used for?

Answer 20

coverts mRNA back into cDNA

Question 21

What is the role of the restriction enzyme in technology?

Answer 21

cuts the DNA into pieces

Question 22

What is meant by “sticky ends”?

Answer 22

a shape of cut that restriction enzymes make that allows new bases to create hydrogen bonds with the old bases