Biotechnology

Question 1

Gel electrophoresis

Answer 1

Separate DNA fragments by size using electric charge
- repels DNA, + attracts DNA
larger fragments slower, smaller fragments faster
Gel is agarose or polyacrylamide
Gel is submersed in buffer to carry electric current
DNA visualized using fluorescent dye

Question 2

Gel electrophoresis blotting

Answer 2

Southern: DNA sample
Northern: RNA a sample
Western: proteins detected using antibodies

Question 3

Polyamerase chain reaction

Answer 3

Amplify small DNA fragment
Kary Mullis
Uses deoxyribonucleotide triphosphate and Rx buffer for enzyme function



Question 4

Polyamerase chain reaction steps

Answer 4

1) Denaturation: high temp
2) Annealing primers: low temp
3) DNA synthesis: medium temp. Taq polymerase

Question 5

What is a primer used in PCR?

Answer 5

Short DNA fragments complementary to both ends of a target sequence

Question 6

What is used in DNA synthesis used in polymerase chain reaction?

Answer 6

The DNA polymerase Taq polymerase isolated from bacteria

Question 7

Polyamerase chain reaction applications

Answer 7

1) multiply minute DNA samples to be analyzed
2) forensics ex. blood and hair follicle cells
3) detection of genetic defects in embryo using a small single cell sample
4) Analysis of mitochondrial DNA

Question 8

Clone

Answer 8

Genetically identical copy

Question 9

Protein coding/Gene coding

Answer 9

Isolation of specific DNA sequence

Question 10

Vector

Answer 10

Carries DNA in host and can replicate
Ex. Bacteria, virus, or yeast

Question 11

Most versatile and common host

Answer 11

E. coli bacteria

Question 12

Small insert size used for cloning small DNA fragments

Answer 12

Plasmids
Small circular chromosomes

Question 13

Large chromosomes with larger insert size for cloning larger DNA segments

Answer 13

Yeast artificial chromosomes
Bacterial artificial chromosomes

Question 14

DNA library

Answer 14

DNA in a vector represents complex DNA mixture

Question 15

Genomic library

Answer 15

Representation of entire genome in vector
1) Genome is randomly fragmented
2) inserted into vector
3) introduced into host cell
4) Constructed in bacterial artificial chromosome

Question 16

Complementary DNA

Answer 16

cDNA
DNA copies of mRNA
1) mRNA isolated
2) reverse transcriptase makes cDNA
3) cDNA used to make library 

Question 17

All genome libraries from one cell are the same, but cDNA libraries from one cell are what?

Answer 17

Different

Question 18

Reproductive cloning

Answer 18

Dolly the sheep what is the first vertebrate mammal cloned
Dying young due to aged DNA


Question 19

Medically important proteins

Answer 19

Can be produced in bacteria, but it’s difficult to purify desired proteins from other bacterial proteins
ex. Human insulin, interferon, arterial peptides, tissue plasminogen activator, and human growth hormone

Question 20

Vaccines

Answer 20

Subunit vaccines: genes in coding protein coat spliced into genome fragment.
DNA vaccines: Depend on cellular immune response (not antibodies)

Question 21

Gene therapy

Answer 21

Add functional copy of gene to correct hereditary disorder
Some successes and failures
ex. Severe combined immunodeficiency disease gene therapy gave some children leukemia

Question 22

Transgenic animals

Answer 22

Contain recombinant DNA
used to express human genome
ex. Mice, sheep, and goats

Question 23

Transgenic plants

Answer 23

Genetically modified organics
Genes resistant to disease herbicides and pesticides
ex. Bacterial toxin (bt toxin): Natural insecticide Gene
Flavr savr tomato

Question 24

Study of genomes a DNA level

Answer 24

Genomics

Question 25

Study of entire set of proteins

Answer 25

Proteomics

Question 26

Study of small molecules found in organisms 

Answer 26

Metabolomics

Question 27

Study of entire mRNA set expressed in an organism

Answer 27

Transcriptomics

Question 28

Study of genetic material maternally inherited next generation

Answer 28

Mitochondrial genomics

Question 29

Study of collective genomes within an environmental sample

Answer 29

Metagenomics

Question 30

Study of transcription of genes in response to a chemical or drug

Answer 30

Pharmacogenomics Toxicogenomics

Question 31

J. Craig Venter

Answer 31

First to sequence human genome First to generate alternate life form

Question 32

CRISPR

Answer 32

Clustered regularly interspaced short palindromic repeats Synthesize guide RNA (gRNA) for desired sequence, like repaired gene Insert gRNA and CRISPR CAS 9 Will re-synthesize the complementary DNA to the gRNA Allows you to answer any sequence into germline DNA of the cell Adaptive immune mechanism for bacteria to fight bacteria phage viruses by identifying similar viral sequences from a previous infection

Question 33

Autologous priming and replacement

Answer 33

1) remove DC/T cells from patient 2) Prime cells with cytokines and tumor antigens 3) Put cells back in patient