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 GEL ELECTROPHORESIS OF DNA ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science APPLICATIONS OF GENETIC ENGINEERING: DNA CLONING Genetically engineered bacteria Genetic engineering relies on DNA cloning Process of producing copies of DNA Cloned DNA generally combined with carrier molecules called cloning vector or plasmid Independently replicating DNA molecule Insures replication of target DNA (Insert) Recombinant molecule ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science TECHNIQUES USED IN GENETIC ENGINEERING Obtaining DNA to be cloned Generating a recombinant molecule Restriction enzymes and ligases used to create recombinant molecule Introducing recombinant DNA into new host Host acts as “incubator” for DNA replication DNA-mediated transformation often used to get DNA into host ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science OBTAINING DNA TO BE CLONED Genomic library containing either natural copies of genes cDNA library containing copies of genes made from mRNA Synthetic DNA ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science CONSTRUCTION OF GENOMIC LIBRARIES Used when gene of interest is on a chromosome that has not been sequenced The library is constructed by cleaving the genome and then cloning the fragments into vectors The libraries are screened for the genes of interest in a variety of ways ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science GENERATING A GENOMIC DNA LIBRARY Each cell contains one fragment of a given organism’s genome. ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science cDNA LIBRARY: MAKING cDNA FROM EUKARYOTIC mRNA ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science SYNTHETIC DNA Under certain circumstances, genes can be made in vitro using a DNA synthesis machine Oligonucleotides (can be DNA or RNA) is made by a DNA synthesis machine The sequence of the gene must be known before it can be synthesized Synthesized in stepwise process in which single nucleotides are added to end of a growing chain Chain length can vary from short (2-30 nucleotides) to long (50-100 nucleotides) Can be synthesized to have known nucleotide sequences Primers for cloning and PCR ISBN: 978-0-8153-6514-3...
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