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TORTORA • FUNKE • CASE Microbiology AN INTRODUCTION EIGHTH EDITION B.E Pruitt & Jane J. Stein Chapter 9, part A Biotechnology and Recombinant DNA PowerPoint® Lecture Slide Presentation prepared by Christine L. Case Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Biotechnology and Recombinant DNA • Biotechnology: • The use of microorganisms, cells, or cell components to make a product • Foods, antibiotics, vitamins, enzymes • Recombinant DNA Technology: • Insertion or modification of genes to produce desired proteins Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.1.1 Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.1.2 Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Table 9.1.1 Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Table 9.1.2 Selection & Mutation • Selection: Culture a naturally-occurring microbe that produces desired product • Mutation: Mutagens cause mutations that might result in a microbe with a desirable trait • Site-directed mutagenesis: Change a specific DNA code to change a protein • Select and culture microbe with the desired mutation Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Restriction Enzymes • Cut specific sequences of DNA • Destroy bacteriophage DNA in bacterial cells • Cannot digest (host) DNA with methylated cytosines Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.2 Vectors • Carry new DNA to desired cell • Shuttle vectors can exist in several different species • Plasmids and viruses can be used as vectors Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Vectors Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.3 Polymerase Chain Reaction (PCR) • To make multiple copies of a piece of DNA enzymatically • Used to • Clone DNA for recombination • Amplify DNA to detectable levels • Sequence DNA • Diagnose genetic disease • Detect pathogens Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings PCR Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.4.1 PCR Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.4.2 DNA can be inserted into a cell by: • Transformation • Electroporation • Protoplast fusion Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.5b DNA can be inserted into a cell by: • Microinjection • Gene gun Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings Figure 9.6 & 7 Obtaining DNA • Gene libraries are made of pieces of an entire genome stored in plasmids or phages • cDNA is made from mRNA by reverse transcriptase • Synthetic DNA is made by a DNA synthesis machine Copyright © 2004 Pearson Education, Inc., publishing as Benjamin Cummings
