feb 24 notes

feb 24 notes - Anatasia Gangadin Cell biology Today...

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Anatasia Gangadin Page 1 10/6/2009 Cell biology Today: Molecular biology techniques Find a copy of the book for test *Next week: exam 1, 3/3/09 Mutations Gene function o Yeast is a great organism to work with Eukaryotic REALLY easy to grow Bit unusual because either can be haploids or diploids Temperature sensitive mutants Easy to mutate Add mutagen, plate yeast, make replica plates, different temps. Yeast grow at 23* Different types of mutations o Suppressor Rare, unusual Wild type yeast, gene A and B. One gene is mutant, therefore, phenotyope is not normal. o Synthetic Lethality 1 Focuses on redundant pathways o Synthetic Lethality 2 Temperature sensitive mutants o Reference for 2 different temperatures. 23 centigrade is permissible temperature 36 centigrade is non permissible temperature Heat sensitive protein that denatures at 36*. o Complementation analysis If 2 genes are related to same function Begin with 2 haploid mating types of yeast, Each one has a mutation with cell cycle, but not sure where the mutation is. Mated, forms diploids, temperature sensitive mutant. Since both grow at temp sensitive, the mutation is on different alleles General principles and concepts Restriction nucleases o Beginning of modern molecular biology techniques 1970’s. Before then, you could purify DNA, but you couldn’t manage DNA. It was just long stringy mess. After 1970’s, restriction nucleases were discovered and could cut DNA. Length is managed, less complicated. First discovered in Bacteria
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Anatasia Gangadin Page 2 10/6/2009 Cell biology To deal with invading viruses; cut up virus and virus DNA Commercially available o Can cut with blunt ends or sticky ends (preferred) Gene splicing and transfection o Many procedures o Plasmid cloning vector o DNA Ligase= knits Nucleic acid Hybridization o Complimentary nature of DNA and RNA Polymerase chain reaction o Cloning DNA engineering o Considered where you alter the gene (Transfected) for some purpose o Genetic Herceptin Not hybridomas, but Tiny Hamster ovary cells
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feb 24 notes - Anatasia Gangadin Cell biology Today...

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