Construction of p22 challenge phage, experiment 8.

Overview: Bacterial Pathogenesis: General concepts in the strattera no prescritpion pathogenesis of bacterial, Bacterial Pathogenesis paradigm: Salmonella Typhimurium, Vibrio cholerae

Practical genetics of microorganisms: Genetic nomenclature; microbiological procedures, culture media, antibiotics, resistance to antibiotics and positive and negative selection, bacterial physiology

genetic mapping: Mutants and Analysis: Mergers and transposons: The use of transposons in bacterial genetics bacteriophage Mu; operon and Jean M: operon and gene fusion with LacZ, operon fusion with the uidA (GUS ) Gene fusion with the PhoA, in vivo expression technology

gene regulation: recombinant DNA experiments: introduction to experiments 16; Experiment 1. Transposons: Mini-Mu-lac

insertion in the structural genes, Experiment 2. Transposons: T 10 A related insertion of structural genes, Experiment 3. Transposons: Tn 10 inserts in regulatory genes, Experiment 4. Legend (sensitive to heat and yellow) mutation, Experiment 5. In Vivo Molecular Cloning; Experiment 6. Physical mapping of bacterial chromosomes;

Introduction to experiments 711, Experiment 7. Construction of phage P22 problem, Experiment 8. Challenge phage tests, experiment 9. Selecting an operator mutation, Experiment 10. DNA sequence analysis of mutant phage challenge, Experiment 11. Select the second site suppression mutations that recognize mutant operator sites;

Introduction to the 1215 experiments, Experiment 12. Select Vibrio cholerae T insertion PhoA; Experiment 13. Southern hybridization of DNA to the map Tp PhoA insertion, Experiment 14. Allelic exchange in Gram-negative bacteria use suicide plasmid vectors, Experiment 15. Oligonucleotide directed site-specific mutagenesis

Programs strains, phages and plasmids:

Lists deformation strains of bacteria, bacteriophages, plasmids

Culture Media and Supplements: Liquid culture media, solid growth media, culture media supplements , antibiotics, buffers, P22 media indicator, MOPS culture medium

collection strain: strains storage, delivery strains, strains of the collection of registration letter

Genetic exchange and reflection: generalized transduction phage P22, a quick reflection in S. Typhimurium with Mud-P22 profahov, phage mini-lysates, hydroxylamine mutagenesis of plasmid DNA, preparation and transformation of competent cells, preparation of electrocompetent cells and electroporation

analysis of enzymes: galactosidase analysis in permeable cells, alkaline phosphatase in the analysis of permeable cells

recombinant DNA methods: plasmid minipreps DNA restriction endonuclease buffers, ethanol precipitation, phenol extraction, Spin columns, Drop dialysis, agarose gels, nedenaturiruyuschih electrophoresis in polyacrylamide gels of DNA, DNA molecular weight standards useful, purification and quantitative PCR products, purification of DNA groups from agarose gel by QIAquick extraction, Southern hybridization solution, display fusion joints by combining the Southern and Western analysis of oligonucleotide directed site-specific mutagenesis

plasmids and transposons card restrictions: plasmid prevaricate-1, plasmids pEG5005, plasmid and pSU18 pSU19, plasmids pGP704, plasmids and pKAS32 pKAS46, plasmids pPY190, plasmid pPC36, plasmid pMS421, plasmids pGW1700, bacteriophage Mu derivatives, transposons Tn 10 derivatives transposons T PhoA

link