Chapter 8 Recombinant DNA Technology

Question 1

Biotechnology

Answer 1

the use of microorganisms to make practical products

Question 2

Recombinant DNA Technology

Answer 2

intentionally modifying genomes of organisms for practical purposes

Question 3

3 goals of Recombinant DNA Technology

Answer 3

1 eliminate undesirable phenotypic traits
2 combine beneficial traits of 2 or more organisms
3 create organisms that synthesize products humans needs

Question 4

5 tools of Recombinant DNA Tech

Answer 4

1 mutagens
2 reverse transcriptase
3 restriction enzymes
4 vectors
5 gene libraries

Question 5

Mutagens

Answer 5

-physical + chemical agents that produce mutations in DNA

Question 6

Mutagens for Recombinant DNA

Answer 6

• used to create changes in microbe’s genomes to change phenotype
• selects for + cultures cells w beneficial characteristics
• mutated genes alone can be isolated

Question 7

Reverse Transcriptase

Answer 7

an enzyme that uses RNA template to transcribe a molecule of cDNA

(turns RNA to DNA)

Question 8

Reverse Transcriptase for Recombinant DNA

Answer 8

• isolated enzymes fr retrovirus
• easier to isolate mRNA for desired protein
• cDNA generated from mRNA of eukaryotes has intron removed to allow prokaryotic cells to produce eukaryotic proteins

Question 9

removal of introns from cDNA from eukaryotic mRNA. This allows for…..

Answer 9

prokaryotic cells to produce eukaryotic proteins

Question 10

cDNA

Answer 10

complimentary DNA

Question 11

Restriction Enzymes/Endonucleases

Answer 11

• bacterial enzymes that cut DNA molecs only at restriction sites
• creates recombinant DNA by joining fragments

Question 12

2 types of Restriction Enzymes/Endonucleases

Answer 12

1 cuts w sticky ends
2 cuts w blunt ends

-categories are based on type of cut

Question 13

Vectors

not marked

Answer 13

nucleic acid molecules that deliver a gene into a cell

-includes viral genomes, transposons, and plasmids

Question 14

Gene Libraries

not marked

Answer 14

• collection of bacterial or phage clones
• each clone in library often contains one gene of an organism’s genome
• provides a ready source of genetic material

Question 15

Polymerase Chain Reaction [PCR]

Answer 15

• multiplying DNA in vitro (like a xerox machine)
• large number of molecs of DNA are produced
• can be automated using a thermocycler

Question 16

3 repetitive steps of PCR

Answer 16

1 denaturation
2 priming
3 extension

Question 17

why is it important to be able to amplify DNA?

Answer 17

• epidemiologists use to amplify genome of unknown pathogen

- amplified DNA from Bacillus anthraces spores in 2001 to identify source of spores

Question 18

2 methods of inserting DNA into cells

not marked

Answer 18

1 Natural Method

2 Artificial Method

Question 19

Gene Gun + Microinjection

not marked

Answer 19

-artificial method of injections

Question 20

Gene Mapping

not marked

Answer 20

• locating genes on a nucleic acid molecule

- provides useful facts concerning metabolism, growth characteristics, and relatedness to others

Question 21

Nucleotide Sequencing

Answer 21

• used to relate DNA sequence to protein functions
• Genomics
• elucidation of the genomes of pathogens is a priority

Question 22

Genomics

Answer 22

sequencing and analysis of the nucleotide bases of genomes

Question 23

Protein Synthesis

not marked

Answer 23

creation of synthetic proteins by bacteria and yeast cells

Question 24

Vaccines

not marked

Answer 24

• production of safe vaccines

- subunit vaccines

Question 25

new approaches to stimulate immunological memory ***not marked***

Answer 25

- intro of genes of pathogens into fruits + veggies - injecting humans w plasmid carrying gene fr pathogen - --human synth pathogen's proteins

Question 26

Gene Screening ***not marked***

Answer 26

- DNA microarrays are used to screen individuals for inherited disease caused by mutations - can also ID pathogen's DNA in blood or tissues

Question 27

Gene Therapy

Answer 27

- missing or defective genes are replaced w normal copies - some patient's immune system react negatively - treatment of cancer by directing toxin-loaded virus to the cancer cells

Question 28

DNA fingerprinting

Answer 28

ID individuals or organisms by their unique DNA sequence

Question 29

Gene Therapy for killing cancer ***not marked***

Answer 29

TERT gene product> Telomerase | -hTERT promoter is active in all types of tumors and immortal cells but inactive in somatic tissues

Question 30

Telomerase

Answer 30

- highly active enzymes that divide rapidly in cells | - abnormally active in most cancer cells which grow + divide w/o control or order

Question 31

Sticky ends

Answer 31

able to pair w other complimentary ends

Question 32

Blunt ends

Answer 32

ligase brings blunt ends together

Question 33

E.Coli DNA polymerase vs. Thermus aquaticus (TAQ) DNA polymerase

Answer 33

- TAQ DNA polymerase is able to withstand protein-denaturing conditions (high heat), whereas E.Coli is not - E.Coli was originally used for PCR but has been replaced w TAQ - used for DNA aplification

Question 34

Gel Electrophoresis

Answer 34

uses electrical charge to separate molecules according to their size -separates DNA fragments by size

Question 35

reverse transcriptase is found in which organism ?

Answer 35

certain RETROVIRUSES like HIV virus