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

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Answer 13

nucleic acid molecules that deliver a gene into a cell

-includes viral genomes, transposons, and plasmids

Question 14

Gene Libraries

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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

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Answer 18

1 Natural Method

2 Artificial Method

Question 19

Gene Gun + Microinjection

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Answer 19

-artificial method of injections

Question 20

Gene Mapping

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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

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Answer 23

creation of synthetic proteins by bacteria and yeast cells

Question 24

Vaccines

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Answer 24

• production of safe vaccines

- subunit vaccines

Question 25

new approaches to stimulate immunological memory

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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

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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

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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