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26 Cards in this Set

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  • Back
What are restriction endonucleases?
They are tools used in genetic engineering that recongize a restriction site on DNA and cut it.
What are the types of restriction endonucleases? What is different about them? Which is used more in genetic engineering?
Type 1- cuts straight through both DNA strands. Type 2- cuts with dyad symetry leaving a sticky end to the DNA. Type 2 is used more than Type 1.
What do genetic engineers use to attach the sticky ends of DNA fragments together?
DNA Ligase.
One of the ways that genetic engineers mass produce recombinant DNA is by using cells as factories in a _____/________ System.
What is the most common host in a Host/Vector System? What are the most common vectors?
E. Coli. Plasmids and Phages.
What are plasmids? What are phages?
Small, circular, extrachromasomal pieces of DNA that are dispensible to the cell. Viruses that infect bacteria.
What are the two things that a plasmid must have?
They need an origin of replication and a marker that allows antibiotic resistance.
Where is the circular plasmid inserted so that it is cut into a linear plasmid?
A Multiple Cloning Site.
How is the presence of recombinant DNA detected?
What virus phage are most of the phages in genetic engineering based on?
What is necessary in order to use a phage as a vector?
There must be live cells to replicate and the recombinant DNA must be in the phage's head.
What is recombinant DNA?
It is DNA that has been engineered in a lab from DNA fragments from different genomes.
What is a DNA library? What is a genomic library?
A collection of DNA that can be inserted into a host. The entire genome in a vector.
What are cDNA libraries and how are they acquired?
cDNA libraries are collections of expressed DNA. They are formed by using reverse transcriptase, isolated from retroviruses, to convert mRNA to DNA.
What are the four stages of the genetic engineering expirement?
1. DNA Cleavage, 2. Production of Recombinant DNA, 3. Cloning, 4. Screening
After cleaving DNA with endonucleases, how are the fragments separated?
What are the two steps of screening?
Primary Screening- eliminate hosts that contain no vector or contain an empty vector. Secondary Screening- hosts with the gene of interest isolated.
How is secondary screening carried out?
Hybridization- a probe nucleic acid (usually radioactive) binds to the desired gene.
What is a Polymerase Chain Reaction? What are its steps?
A process for producing many copies of a DNA fragment. Denaturation, Annealing of Primers, Primer Extension.
What process is used to identify a DNA fragment?
Southern Blotting- DNA from gel electrophoresis tranfered to nitrocellulose. Primer poured on nitrocellulose.
What are Restriction Fragment Length Polymorphisms (RFLPs) used for?
They can be used to identify an individual from a sample of their genes.
How is gel electrophoresis used in criminal investigations?
DNA Fingerprinting.
What are some of the medical applications of genetic engineering?
Pharmaceuticals like artrial peptides to handle high blood pressure and kidney failure, tissue plasminogen activators to dissolve blood clots, gene therapy, and piggyback vaccines.
What are some of the agricultural applications of genetic engineering?
Nitrogen fixation, herbicide resistance, and insect resistance, genetically modified plants.
What plasmid has been used in agricultural genetic engineering?
The TI Plasmid.
Who made these flashcards?
Robert Fromm