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BIOL 200 Study Guide - Final Guide: Transcription Preinitiation Complex, Transcription Factor Ii E, Stamen

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School
McGill University
Department
Biology (Sci)
Course
BIOL 200
Professor
Mathieu Roy

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Document Summary

Bacterial restriction enzymes cut dna at specific sites. Dna can be amplified from this vector. Studying genes of interest: dna libraries: digest entire genomes, first: mechanically, second: restriction enzyme digests, sequences are kept in genomic libraries, one genome fragment per plasmid cdna libraries - represent mrna tissue specificity, abundance. Specialized vectors permit efficient expression in higher eukaryotic cells: transient transfection transfect cultured cells with a vector. Protein is expressed from cdna in plasmid dna: not all cells will take up the plasma, eventually all cells will get rid of the plasmid. No centromere (no faithful segregation) => loose plasmid: stable transfection (do not use the term transformation) vector contains a antibiotic resistance gene; therefore only cells which contain the vector will survive. Whenever cells lose the vector they die: when the vector is transfected into the host chromosome => becomes stable.

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Related Questions

1. What are essential elements for a useful plasmid vector? Why is it advantageous to have a restriction site for an endonuclease in a plasmid vector? Can a piece of foreign DNA be inserted into a plasmid if they have been cut with different restriction endonucleases?

2. Briefly explain the molecular basis of cDNA synthesis. Why is cDNA synthesis considered to be such an important technique in molecular biology?

Why is it important that Type II restriction endonucleases recognition sequences located with the multiple cloning site (MCS) not occur anywhere else within a plasmid?

A-Which of the following is NOT one of the steps required toclone a piece of DNA using a plasmid vector?
1. Cut a vector and a DNA insert with aspecific restriction enzyme to create complementary "stickyends."
2. Isolate DNA fragments by gelelectrophoresis.
3. Ligate insert DNA and a vector withsimilar sticky ends together.
4. Transform ligated pieces of DNA into E.coli.
5. Select the plasmid vector that hasreplaced its origin of replication with insert DNA.

B-A representative genomic library of anorganism's DNA fragments results from the use of
1. DNA ligase joining multiple types ofvectors.
2. partial digestion with frequentlycutting restriction endonucleases.
3. many pieces of recombinant DNA fromseveral different types of organisms.
4. viruses that cleave the DNA into randompieces.

C-One of the most usefulmethods for identifying a specific DNA sequence is
1. the Southern blot.
2. the Western blot.
3. magnetic resonance imaging.
4. the Eastern blot.
5. thin-layerchromatography.