JBiotech - DNA Technology DNA T E CHNO L O G Y But first...

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DNA Technology Heyer 1 DNA TECHNOLOGY But first before we splice a gene Lets think about the unforeseen Our good intent to hybridize Could engineer a bad surprise Similar terms — often used interchangeably Biotechnology: the manipulation of organisms to produce a product. Fermentation, artificial breeding, pharmaceutical and nutritional supplements, and now… Genetic engineering: the direct manipulation of an organism’s DNA. Recombinant DNA: insertion of DNA from one source into another. Transgenics: producing an organism with foreign DNA inserted into its genome. TERMINOLOGY TERMINOLOGY Restriction Digests RFLP — “genetic fingerprinting” Hybridizations & Molecular Probes Polymerase Chain Reaction (PCR) Recombinant DNA Gene Cloning Transgenics Gene Expression Analyses Genome Mapping & Sequencing TOOLS & TECHNIQUES RESTRICTION ENZYMES RESTRICTION ENZYMES • Bacteria produce special enzymes to chop up viral DNA. • Biotechnologist use these “restriction enzymes” to cut DNA in specific places (restriction sites). • Many restriction enzymes cut the DNA polymer in a staggered pattern that produce “sticky” single-stranded ends to the DNA fragments. RESTRICTION ENZYMES DNA Technology as a diagnostic tool Everyone’s DNA is unique Closer the relationship the more similar the DNA R estriction F ragment L ength P olymorphisms RFLPs “Ruff-lips” RESTRICTION FRAGMENT ANALYSIS RESTRICTION FRAGMENT RESTRICTION FRAGMENT
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DNA Technology Heyer 2 RESTRICTION DIGEST RESTRICTION FRAGMENTS RESTRICTION DIGEST RESTRICTION DIGEST RESTRICTION FRAGMENTS Electrophoresis of fragments RESTRICTION FRAGMENT LENGTH POLYMORPHISM RESTRICTION FRAGMENT LENGTH POLYMORPHISM Criminology USES OF RESTRICTION FRAGMENT ANALYSIS USES OF RESTRICTION FRAGMENT ANALYSIS Missing persons USES OF RESTRICTION FRAGMENT ANALYSIS USES OF RESTRICTION FRAGMENT ANALYSIS Criminology Missing persons Paternity USES OF RESTRICTION FRAGMENT ANALYSIS RESTRICTION FRAGMENT ANALYSIS Medicine I nborn errors of metabolism Carriers Prenatal testing Provirus DNA USES OF RESTRICTION FRAGMENT ANALYSIS USES OF RESTRICTION FRAGMENT ANALYSIS
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DNA Technology Heyer 3 Genetic markers RESTRICTION FRAGMENT ANALYSIS ANALYSIS MOLECULAR PROBES CAN IDENTIFY SPECIFIC GENES MOLECULAR PROBES CAN IDENTIFY SPECIFIC GENES Hybridization: DNA/RNA hybrid molecule Southern Blotting RESTRICTION FRAGMENT ANALYSIS RESTRICTION FRAGMENT RESTRICTION FRAGMENT ANALYSIS RESTRICTION FRAGMENT Comparing two different DNA molecules, such as two alleles for a gene Figure 20.9a,b Normal b -globin allele Sickle-cell mutant b -globin allele 175 bp 201 bp Large fragment Dde I Dde I Dde I Dde I Dde I Dde I Dde I 376 bp Large fragment Dde I restriction sites in normal and sickle-cell alleles of b -globin gene.
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