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24 Cards in this Set

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What is RNA interference?
Turns off expression of defective genes. It does this through short interfering RNA’s to create double stranded mRNA. Thus the end result is cell views it as double stranded RNA which is foreign to the cell and destroys it.
The human genome project did 3 things?
Found nucleotide sequences for all the genes in the body or most
Helps to provide diagnostics and treatments
Used random shotgun sequencing used to sequence the genome very rapidly.
What is the process for which random shotgun sequencing works?
Isolate DNA
Fragment DNA with restriction enzymes
Clone DNA in a bacertial artificial chromosome (BAC)
Sequence DNA fragments
Assemble sequences
Edit sequences fill in gaps
it is a 6 step process
Insulin supplement used to come from animals now where does it come from ______________.
Answer: E. coli
Gamma-interferon is the treatment of chronic granulomatous diesease what microorganism which now made by E. coli because of biotechnology
Answer: True
This is a product of genetic engineering and corrects growth deficiencies in children, it is also produced by E. coli ________ ___________ __________ .
Answer: Human Growth hormone.
How do we clone DNA?
Vector such as a plasmid is isolated
DNA is cleaved by an enzyme into fragments
Gene is inserted into plasmid
Plasmid is taken up by a cell such as a bacterium
Cells with gene of interest are cloned
(Clue: It is a 5 step process.)
In most applications of genetic engineering you either want the gene or its product called a _____________.
Answer: protein.
What do restriction enzymes do?
They cut specific sequences of DNA. They exist in bacteria as a defense mechanism against phages. Bacteria methlyases it own genome to prevent self digestion
How does restriction enzymes cut DNA?
They cut DNA @ specific sequences with sticky ends.
Process of altering DNA:
Digest DNA plasmid & DNA of interest
Combine in tub-DNA linesup
DNA ligase joins together
3 step process
What do shuttle vectors do?
They carry new DNA to desired cell.
How is PCR done?
Heat to 94C
Add primers
Primers attach @ 60C
Incubate @ 72 C making copies
The cycle is repeated to make more copies
5 step process
When we know little about the gene sequence what can we use?
Genetic Library, which are made of pieces of an entire genome stored in plasmids or phages.
How do we do this?
Restriction enzymes & Cloning
This process is a great way to obtain all of ------------ from an organism.
genetic material
What are two challenges that people face when they are trying to make a genetic library?
Bacteria do not have introns & extons (We want only extons fully spliced) & Tissue type experiments, are done better with CDNA library
How do we get DNA into cells?
Transformation, electroportation, protoplast fusion.
3 ways
3 more ways to insert DNA.
-gene gun
How do we use a selection?
Culture a naturally-occuring microbe that produces desired product
How do we use a mutation?
Mutagens cause mutations that might result in a microbe with a desirable trait
How do we use site directed mutagenesis?
Change a specific DNA sequence to change a protein
Cloning vector and 4 different ways to get DNA to put in that vector.
1. PCR-specific DNA seq. 2. genome-digest w restriction enzymes-ligate in to create genetic library. 3. mRNA sDNA Digested w/restriction enzymes or inserted directly
4. Make DNA 1 base at a time-be purchased
How do we know which bacterium has DNA of interest?
We know a bacterium has DNA inserted into a vector via B-galactosidase blue white assay.