Genetic Engineering - Microbiology - Lecture Slides, Slides of Microbiology

Download Genetic Engineering - Microbiology - Lecture Slides and more Slides Microbiology in PDF only on Docsity! DNA TECHNOLOGY Genetic engineering and all that stuff Docsity.com Glowing mice Docsity.com Overview of recombinant DNA technology Insert plasmid and gene into bacterium. O O \ 6 Culture bacteria. Harvest copies of gene to insert into plants or animals Harvest proteins coded by gene = | Eliminate undesirable phenotypic traits Create beneficial combination of traits Produce vaccines, antibiotics, hormones, or enzymes FIGUBBSRahyZoom Plasmid Vectors Example plasmid (pBR322) Tetracycline Resistance Ampicillin Bam H1 Resistance Pru | Sal 1 Pst 1 Origin of Replication ® Docsity.com Tools and Techniques • Restriction Enzymes Cut DNA at precise locations based upon a specific target sequence of nucleotides • DNA Ligase Enzyme that “seals” or ligates DNA pieces together Docsity.com Cloning Steps • Isolate gene of interest (cut out of genome with a Restriction Enzyme) • Cut Vector with same R.E. to get compatible ends • Mix 2 DNAs together and Ligate (DNA Ligase) • Transform • Screen for successful event • Celebrate Docsity.com Cloning a human gene in a bacterial plasmid: a closer look Docsity.com Inserting DNA into Cells • Goal of DNA technology is insertion of DNA into cell (transformation) • Can use vectors and natural methods of transformation, transduction, and conjugation • Artificial methods – Electroporation – Protoplast fusion – Injection – gene gun and microinjection Docsity.com Tools and Techniques • GEL ELECTrOpHOrEsis Docsity.com Gel Electrophoresis Wells (-) Electrophoresis ; chamber filled with ; buffer solution Agarose gel (+) ) @& Movement of DNA Xe Lane of DNA fragments of known sizes (kilobase pairs) ® Docsity.com Gel Electrophoresis of Macromolecules —s«~ f = .. \ —<— Docsity.com DNA fingerprints from a murder case Defendant’s i | - blood = o a Blood from j h ; | | ! i Ri tae defendant’s = clothes it OD 4 ali i § ' 1 eet: < Victim’s blood ‘Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. ® Docsity.com DNA Fingerprinting FiGurE 8.13 M = MOTHEr F = FATHEr C = CHiLDrEN both children show some bands with each parent Docsity.com Tools and Techniques • DNA sYNTHEsis • DNA sEquENCiNG Docsity.com n — wo oO c o 5 oa M n <x 2 a Tools and Techniques • pCr (pOLYMErAsE CHAiN rEACTiON)  Large number of identical molecules of DNA are produced in vitro  Allows any piece of DNA to be rapidly amplified !  Only minute amounts of DNA are needed ! Docsity.com > Animation: Polymerase Chain Reaction STARTING MATERIALS: DNA polymerase Primers: ie an @ Heat to separate DNA strands © Coolto allow primers to © DNA polymerase extends the 3’ end of each primer / | vt WELT TT ‘Copyright @ Pearson Education, Inc., publishing as Benjamin Cummings. DNA 5 7 Targeted sequence Y XN i ~ i i Nucleotides: dATP acTP dGTP dTTP Cyc.e 1 yields 2 molecules CYcLe 2 yields 4 molecules Cyrc_e 3 yields 8 molecules Docsity.com AppLiCATiONs • Diagnosis of Disease • Gene Therapy • Vaccines and other Pharmaceuticals • Forensic use • Environmental use (bioremediation) • Agricultural use * Transgenic animals * Transgenic plants Docsity.com Transgenic organisms Docsity.com “Golden” rice contrasted with ordinary rice Transgenic Golden Rice (High levels of Beta-carotene –used to make Vitamin A – give the rice a golden color. Docsity.com Using the Ti plasmid as a vector for genetic engineering in plants TRANSGENIC PLANT Docsity.com Safety and Ethics GMO (genetically modified organism) food controversy Docsity.com \ WONT EAT ANYTHING THATS GENETICALLY IT COULD BE UNHEALTHY. Docsity.com BT Corn basics • BT protein is produced by the bacterium Bacillus thuringiensis. Toxic to some insects, nontoxic to all other life forms. • BT corn has the gene inserted so that the corn plant makes the BT protein. • Ingestion of BT protein by the larvae of the European Corn Borer kills the larvae • The corn plant now has it’s own defense Docsity.com Pluses and minuses • Reduced need for pesticides • Greater Yield, etc. etc. • Potential impact on other species None of any consequence noted…. …but still the GREAT UNKNOWN. Docsity.com