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Recombinant DNA (rDNA) technology: Insertion or modification of genes to produce desired proteins
Figure 9.1
Figure 9.1
Table 9.2
Table 9.2
Restriction Enzymes
Cut specific sequences of DNA Destroy bacteriophage DNA in bacterial cells
Table 9.1
Figure 9.2
Vectors
Carry new DNA to desired cell Plasmids and viruses can be used as vectors
Figure 9.3
PCR
Figure 9.4
PCR
Figure 9.4
PCR
Figure 9.4
Figure 9.5b
Figure 9.5a
Transformation
A Gene Gun
Figure 9.6
Figure 9.7
Obtaining DNA
Genomic libraries are made of pieces of an entire genome stored in plasmids or phages
Figure 9.8
Obtaining DNA
Complementary DNA (cDNA) is made from mRNA by reverse transcriptase
Figure 9.9
Obtaining DNA
Synthetic DNA is made by a DNA synthesis machine
Figure 9.10
Selecting a Clone
Figure 9.11
Copyright 2010 Pearson Education, Inc.
Selecting a Clone
Figure 9.11
Selecting a Clone
Figure 9.12
Copyright 2010 Pearson Education, Inc.
Selecting a Clone
Figure 9.12
Copyright 2010 Pearson Education, Inc.
Making a Product
E. coli Used because it is easily grown and its genomics are known Need to eliminate endotoxin from products Cells must be lysed to get product
Figure 9.13
Making a Product
Saccharomyces Mammalian cells cerevisiae May express Used because it is easily eukaryotic genes grown and its genomics easily are known Harder to grow May express eukaryotic genes easily Plant cells and whole plants May express eukaryotic genes easily Plants easily grown
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Therapeutic Applications
Human enzymes and other proteins Subunit vaccines Nonpathogenic viruses carrying genes for pathogen's antigens as DNA vaccines Gene therapy to replace defective or missing genes
Scientific Applications
Understanding DNA Sequencing organisms' genomes DNA fingerprinting for identification
Figure 9.17
Southern Blotting
Figure 9.16
Southern Blotting
Figure 9.16
Southern Blotting
Figure 9.16
Forensic Microbiology
PCR Primer for a specific organism will cause application if that organism is present Real-time PCR: Newly made DNA tagged with a fluorescent dye; the levels of fluorescence can be measured after every PCR cycle Reverse-transcription (RT-PCR): Reverse transcriptase makes DNA from viral RNA or mRNA