Ch 8 - Recombinant DNA Technology
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95 Terms
What is epigenetics?
Modifications to DNA that turn genes on or off without changing the DNA sequence.
What is an epigenome?
Chemical tags on DNA that regulate gene expression.
What is DNA methylation?
Addition of methyl groups to DNA that typically turns genes off.
What is DNA acetylation?
Addition of acetyl groups that typically turns genes on.
Can epigenetic tags be inherited?
Some can be passed to offspring.
What is biotechnology?
The use of microorganisms to make practical products.
What is recombinant DNA technology?
The modification of genomes of organisms.
What are common tools used in recombinant DNA technology?
Plasmids and restriction enzymes.
What are possible outcomes of recombinant DNA technology?
Remove undesirable traits, combine beneficial traits, or create organisms that make useful products.
What is a plasmid?
A small circular DNA molecule that replicates independently of chromosomal DNA.
What is a vector?
A nucleic acid molecule used to deliver a gene into a cell.
Is a plasmid a vector?
Yes.
What is a recombinant vector?
A vector altered to contain a gene of interest.
What is the first step in creating a recombinant plasmid?
Cut the plasmid with a restriction enzyme.
What enzyme joins DNA fragments together?
DNA ligase.
What is transformation?
Introducing recombinant DNA into competent bacterial cells.
Why are transformed bacteria plated on antibiotic media?
To select bacteria containing the recombinant plasmid.
What are restriction enzymes?
Bacterial enzymes that cut DNA at specific restriction sites.
What are sticky ends?
Restriction enzyme cuts that leave overhanging DNA strands.
What are blunt ends?
Restriction enzyme cuts that leave straight DNA ends.
What disease results from insufficient insulin production?
Diabetes.
What was used before recombinant human insulin?
Bovine and porcine insulin.
Who developed recombinant human insulin?
Genentech.
What was the first genetically engineered drug approved by the FDA?
Humulin.
Why was recombinant insulin advantageous?
Lower cost, high production, fewer adverse reactions.
What does PCR stand for?
Polymerase Chain Reaction.
What is PCR used for?
Amplifying DNA.
Name five uses of PCR.
Pathogen identification, DNA fingerprinting, cloning, genetic testing, paternity testing.
What is the template in PCR?
The DNA being copied.
What are primers in PCR?
Short DNA sequences that start DNA synthesis.
What are dNTPs?
Free nucleotides used to build new DNA strands.
What enzyme is used in PCR?
Taq DNA polymerase.
What happens during PCR denaturation?
DNA strands separate.
What temperature is used for denaturation?
92–94°C.
What happens during PCR priming (annealing)?
Primers bind to DNA.
What temperature is used for priming?
About 55°C.
What happens during PCR extension?
Taq polymerase synthesizes new DNA strands.
What temperature is used for extension?
72°C.
How many PCR cycles are typically performed?
About 30.
What machine performs PCR temperature changes?
Thermocycler.
What is gel electrophoresis?
A technique that separates DNA by size, shape, and charge.
What substance is used to make electrophoresis gels?
Agarose.
Which DNA fragments move farther in a gel?
Smaller fragments.
Why does DNA move in an electric field?
DNA is negatively charged.
Toward which electrode does DNA move?
The positive electrode.
How is fragment size determined in gel electrophoresis?
By comparing movement to standards.
What is DNA fingerprinting?
A method of identifying individuals using DNA patterns.
What are the four steps of DNA fingerprinting?
Isolate DNA, PCR amplification, restriction enzyme digestion, gel electrophoresis.
Why do DNA fingerprints differ among people?
Small genetic variations create different fragment sizes.
What is gene mapping?
Determining the location of genes on chromosomes.
What was the Human Genome Project?
A project that sequenced the human genome.
When was the Human Genome Project completed?
2003.
What is gene therapy?
Replacing defective genes with functional copies.
What is a major challenge of gene therapy?
Getting the gene into enough cells.
What does CRISPR stand for?
Clustered Regularly Interspaced Short Palindromic Repeats.
Where did CRISPR originate?
A bacterial defense system against bacteriophages.
What is guide RNA in CRISPR?
The molecule that directs Cas proteins to a target DNA sequence.
What does Cas9 do?
Cuts DNA at a specific location.
What often happens after Cas9 cuts DNA?
The cell repairs the DNA, often creating mutations.
What is the result of many CRISPR mutations?
Loss of gene function.
How can CRISPR help control malaria?
By introducing malaria resistance into mosquito populations.
Why are microbial community studies important?
Most microbes cannot be cultured in the laboratory.
How are unculturable microbes identified?
Using DNA fingerprints.
What is a transgenic organism?
An organism containing genes from another species.
What is another name for a transgenic organism?
Genetically Modified Organism (GMO).
What causes citrus greening disease?
A bacterium spread by the Asian Citrus Psyllid.
How was citrus greening reduced experimentally?
By inserting a resistance gene from Arabidopsis.
What is herbicide tolerance?
Genetic resistance allowing crops to survive herbicides.
What herbicide is commonly associated with GMO crops?
Glyphosate (Roundup).
What is Bt toxin?
A natural insect-killing toxin from Bacillus thuringiensis.
Why are Bt crops useful?
They resist insect pests.
What is Golden Rice?
Rice genetically modified to produce beta carotene.
What vitamin precursor does Golden Rice produce?
Beta carotene.
Why was Golden Rice developed?
To reduce vitamin A deficiency.
What can severe vitamin A deficiency cause?
Blindness.
Methylation effect on genes?
Turns genes off.
Acetylation effect on genes?
Turns genes on.
Restriction enzymes function?
Cut DNA at specific sequences.
Ligase function?
Joins DNA fragments together.
PCR purpose?
Amplify DNA.
PCR step 1?
Denaturation.
PCR step 2?
Priming (annealing).
PCR step 3?
Extension.
Denaturation temperature?
92–94°C.
Annealing temperature?
~55°C.
Extension temperature?
72°C.
PCR enzyme?
Taq DNA polymerase.
Smaller DNA fragments move ____ in a gel.
Farther/faster.
DNA charge?
Negative.
Human Genome Project completion year?
2003.
CRISPR guide RNA role?
Directs Cas9 to target DNA.
Cas9 role?
Cuts DNA.
Gene therapy purpose?
Replace defective genes.
Golden Rice purpose?
Reduce vitamin A deficiency.
Bt toxin source?
Bacillus thuringiensis.