Question 1

How are genes cloned?

Accepted Answer

Cell containing gene of interest inserted into Bacterium Plasmid, then reinserted into bacteria, which then through binary fission, produces copies of particular gene and produces protein products through particular gene expression

Question 2

HIV

Accepted Answer

2 identical ssRNA 
Has Glycoprotein Envelope
Reverse Transcriptase
Infects T Cells w/ CD4 Receptor
1. T-Cells Don't Function
2.Kills the T-Cells
Treatment: 
1. Azidothymidine (AZT)
Targets RT
2. Protease Inhibitor
Knocks out HIV Proteins
Cockta

Question 3

Ti Plasmid

Accepted Answer

From soil bacterium Agrobacterium Tumefaciens
Use Electropporation to introduce
E.g. Golden Rice, transgenic yellow rice that prevents vitamin A deficiency

Question 4

From soil bacterium Agrobacterium Tumefaciens
Use Electropporation to introduce
E.g. Golden Rice, transgenic yellow rice that prevents vitamin A deficiency

Accepted Answer

in vitro gene manipulated animals 
e.g.sheep with better wool, pig with leaner meat

Question 5

Restriction fragment length polymorphism (RFLP's)

Accepted Answer

Difference in restriction sites on homologous chromosomes due to small nucleotide differences by noncoding DNA
Increased # of markers available for mapping human genome and not limited to genetic variations from disease and differences in protein product

Question 6

Southern Blotting

Accepted Answer

uses gel electrophoresis and nucleic acid hybridization
e.g. able to seperate unaffected, affected, and heterozygous (carriers) of diseases like sickle-cell disease
e.g.able to detect Restriction fragment length polymorphisms (RFLP's)

Question 7

Gel Electrophoresis

Accepted Answer

Useful for Comparing 2 different DNA molecules, even as specific as 2 allele variations
e.g.sickle-cell disease is mutation of 1 single nucleotide that is located within the beta-globin gene

Question 8

Polymerase Chain Reaction (PCR)

Accepted Answer

quicker and more selective
Reaction heated to denature
DNA strands allowed to cool, allowing annealing (hydrogen bonding) of short, ss DNA primers
Heat Stable DNA polymerase extends the primers in the 5'-3' direction
(standard DNA polymerase would be

Question 9

Why use eukaryotic host cells?

Accepted Answer

many eukaryotic proteins will not function unless are modified AFTER translation e.g.addition of carbohydrate or lipid groups

Question 10

Yeast Artificial Chromosome (YACs)

Accepted Answer

Can carry a much longer DNA segment than can a plasmid vector, more likely to contain an entire gene rather than a portion like bacteriophages.
Avoids eukaryotic-prokaryotic incompatibility

Question 11

Expression Vector

Accepted Answer

a cloning vector that contains a highly active prokaryotic promoter just upstream of a restriction site where eukaryotic cell was inserted in the correct reading frame. 
Bacteria will recognize the promoter and proceed to express foreign gene
Used When

Question 12

Nucleic Acid Probe

Accepted Answer

After isolating potential bacteria expressing the desired gene, if we know perhaps the protein the gene of interest encodes or its sequence in the genome of a closely related species, a complimentary molecule of either RNA or DNA can be synthesized and fl

Question 13

Cloning Vector

Accepted Answer

DNA molecule that can carry foreign DNA that can carry foreign DNA into cell and replicate
e.g.Plasmid

Question 14

DNA Ligase

Accepted Answer

used to seal the association of RE manipulated DNA
Catalyzes the formation of covalent bonds that close up e.g.sugar-phosphate backbones

Question 15

Restriction Enzymes

Accepted Answer

Recognize sequences containing 4 to 8 nucleotides
As these sequences occur multiple times, RE will make many  cuts yielding RESTRICTION FRAGMENTS

Genome Project

Biology 106 Final Exam

Question	Answer
How are genes cloned?	Cell containing gene of interest inserted into Bacterium Plasmid, then reinserted into bacteria, which then through binary fission, produces copies of particular gene and produces protein products through particular gene expression
HIV	2 identical ssRNA Has Glycoprotein Envelope Reverse Transcriptase Infects T Cells w/ CD4 Receptor 1. T-Cells Don't Function 2.Kills the T-Cells Treatment: 1. Azidothymidine (AZT) Targets RT 2. Protease Inhibitor Knocks out HIV Proteins Cockta
Ti Plasmid	From soil bacterium Agrobacterium Tumefaciens Use Electropporation to introduce E.g. Golden Rice, transgenic yellow rice that prevents vitamin A deficiency
From soil bacterium Agrobacterium Tumefaciens Use Electropporation to introduce E.g. Golden Rice, transgenic yellow rice that prevents vitamin A deficiency	in vitro gene manipulated animals e.g.sheep with better wool, pig with leaner meat
Restriction fragment length polymorphism (RFLP's)	Difference in restriction sites on homologous chromosomes due to small nucleotide differences by noncoding DNA Increased # of markers available for mapping human genome and not limited to genetic variations from disease and differences in protein product
Southern Blotting	uses gel electrophoresis and nucleic acid hybridization e.g. able to seperate unaffected, affected, and heterozygous (carriers) of diseases like sickle-cell disease e.g.able to detect Restriction fragment length polymorphisms (RFLP's)
Gel Electrophoresis	Useful for Comparing 2 different DNA molecules, even as specific as 2 allele variations e.g.sickle-cell disease is mutation of 1 single nucleotide that is located within the beta-globin gene
Polymerase Chain Reaction (PCR)	quicker and more selective Reaction heated to denature DNA strands allowed to cool, allowing annealing (hydrogen bonding) of short, ss DNA primers Heat Stable DNA polymerase extends the primers in the 5'-3' direction (standard DNA polymerase would be
Why use eukaryotic host cells?	many eukaryotic proteins will not function unless are modified AFTER translation e.g.addition of carbohydrate or lipid groups
Yeast Artificial Chromosome (YACs)	Can carry a much longer DNA segment than can a plasmid vector, more likely to contain an entire gene rather than a portion like bacteriophages. Avoids eukaryotic-prokaryotic incompatibility
Expression Vector	a cloning vector that contains a highly active prokaryotic promoter just upstream of a restriction site where eukaryotic cell was inserted in the correct reading frame. Bacteria will recognize the promoter and proceed to express foreign gene Used When
Nucleic Acid Probe	After isolating potential bacteria expressing the desired gene, if we know perhaps the protein the gene of interest encodes or its sequence in the genome of a closely related species, a complimentary molecule of either RNA or DNA can be synthesized and fl
Cloning Vector	DNA molecule that can carry foreign DNA that can carry foreign DNA into cell and replicate e.g.Plasmid
DNA Ligase	used to seal the association of RE manipulated DNA Catalyzes the formation of covalent bonds that close up e.g.sugar-phosphate backbones
Restriction Enzymes	Recognize sequences containing 4 to 8 nucleotides As these sequences occur multiple times, RE will make many cuts yielding RESTRICTION FRAGMENTS

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