TAMS, Dr. Burleson, Biology, Ch. 16
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- 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.
- Host/Vector.
- 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?
- Screening.
- What virus phage are most of the phages in genetic engineering based on?
- Lambda.
- 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?
- Electrophoresis.
- 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