Medical Technology Department, Faculty of Science, Islamic University-Gaza MB M ICRO B IOLOGY Dr. Abdelraouf A. Elmanama Ph. D Microbiology 2008 Chapter 9 Biotechnology and Recombinant DNA

2008 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

2008 Figure 9.1.1

2008 Figure 9.1.2

2008

Table 9.1.1

2008 Table 9.1.2

2008 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

2008 Cut specific sequences of DNA Destroy bacteriophage DNA in bacterial cells Cannot digest (host) DNA with methylated cytosines Restriction Enzymes

2008 Figure 9.2

2008 Carry new DNA to desired cell Shuttle vectors can exist in several different species Plasmids and viruses can be used as vectors Vectors

2008 Vectors Figure 9.3

2008 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)

2008 PCR Figure 9.4.1

2008 PCR Figure 9.4.2

2008 Transformation Electroporation Protoplast fusion DNA can be inserted into a cell by: Figure 9.5b

2008 Microinjection Gene gun DNA can be inserted into a cell by: Figure 9.6 & 7

2008 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 Obtaining DNA

2008 Genetic Engineering Figure

2008 Genetic Engineering Figure

2008 Making a Gene Product Figure

2008 Making a Gene Product Figure

2008 Used because it is easily grown and its genomics are known Need to eliminate endotoxin from products Cells must be lysed to get product E. coli

2008 Making a Gene Product Figure 9.13

2008 Subunit vaccines Nonpathogenic viruses carrying genes for pathogen's antigens as vaccines Gene therapy to replace defective or missing genes Human Genome Project Nucleotides have been sequenced Human Proteome Project may provide diagnostics and treatments Therapeutic Applications

2008 Random Shotgun Sequencing Figure 9.14

2008 Understanding of DNA Sequencing organisms' genomes DNA fingerprinting for identification Scientific Applications Figure 9.16

2008 Southern Blotting Figure

2008 Southern Blotting Figure

2008 Southern Blotting Figure

2008 Agricultural Applications Table 9.2

2008 Genetic Engineering Using Agrobacterium Figure 9.18

2008 Avoid accidental release Genetically modified crops must be safe for consumption and for the environment Who will have access to an individual's genetic information? Safety Issues and Ethics