Isbn-6514-3 microbiology a clinical approach by tony

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Unformatted text preview: ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science SITE-DIRECTED MUTAGENESIS Specific mutations at specific sites Why? Where? Positions that might stabilize useful proteins How? (Please see next slide.) ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science SITE-DIRECTED MUTAGENESIS Uses small synthetic oligonucleotide to introduce specific mutation into a gene of interest ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science PROTEIN EXPRESSION Production of recombinant proteins: Fusion proteins ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science PROBE TECHNOLOGIES: DNA PROBES DNA probes Used to locate specific nucleotide sequences in DNA or RNA samples Probe is a single-stranded piece of DNA tagged with detectable marker such as a radioactive isotope or a fluorescent dye Because of the label, its presence can be easily determined Probe will hybridize to complementary fragment of interest Applications of probe technologies DNA microarray technology ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science DNA PROBES ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science DNA MICROARRAY TECHNOLOGIES Technique used to study expression of specific genes in DNA samples. Compare healthy and diseased tissue samples Genes expressed during host- pathogen interactions DNA arrays are solid supports with fixed patterns of different single-stranded DNA fragments attached Enable researchers to screen sample for numerous sequences simultaneously ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science THE POLYMERASE CHAIN REACTION (PCR) Enables the rapid synthesis of many copies of a specific DNA fragment from a complex mixture of DNA and other cellular components Reaction mix contains: Primers: short, single-stranded segments of DNA Template DNA, dsDNA containing the region to be amplified (target DNA) Thermostable DNA polymerase such as Taq polymerase, heat-stable DNA polymerase from the thermophile Thermus aquaticus Each of the four nucleotides (dATP, dGTP, dCTP, dTTP) Thermocycler is the instrument used ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science PCR AMPLIFIES SELECTED SEQUENCES Creates millions of copies of given region...
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