Microbiology B.E Pruitt & Jane J. Stein AN INTRODUCTION EIGHTH EDITION TORTORA FUNKE CASE Chapter 9, part A Biotechnology and Recombinant DNA

Biotechnology: –The use of microorganisms, cells, or cell components to make a product –Foods, antibiotics, vitamins, enzymes –Old science - cheese, beer, wine and bread Recombinant DNA Technology: –Insertion or modification of genes to produce desired proteins

Figure 9.1.1

Figure 9.1.2

Table 9.1.1

Table 9.1.2

Tools of Biotechnology Enzymes that do the work: –Restriction enzymes –DNA polymerase –DNA ligase –Reverse Transscriptase Vectors to carry DNA –Plasmids –Viruses DNA sources –Synthetic –Gene libraries Important procedures –PCR –Blotting Southern / northern / western

Cut specific sequences of DNA –Palindromes: Madam I’m Adam; Race car; wow and mom In wild cells they at as a type of disease defense by destroying bacteriophage DNA in bacterial cells Cannot digest (host) DNA with methylated cytosines May cut DNA blunt or with “sticky ends” Tools: Restriction Enzymes

Figure 9.2

Carry new DNA to desired cell Shuttle vectors can exist in several different species Plasmids and viruses can be used as vectors Vectors

Figure 9.3

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 Polymerase Chain Reaction (PCR)

PCR Figure 9.4.1

PCR Figure 9.4.2

Transformation Electroporation Protoplast fusion Microinjection transduction DNA can be inserted into a cell by: Figure 9.5b

Microinjection Gene gun DNA can be inserted into a cell by: Figure 9.6 & 7

Gene libraries are made of pieces of an entire genome stored in plasmids or phages cDNA is made from mRNA by reverse transcriptase –Why??? Synthetic DNA is made by a DNA synthesis machine Obtaining DNA

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 Selection & Mutation