Transcript Document

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
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Table 9.1.1
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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
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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
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Vectors
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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
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PCR
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Figure 9.4.1
PCR
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Figure 9.4.2
DNA can be inserted into a cell by:
• Transformation
• Electroporation
• Protoplast fusion
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Figure 9.5b
DNA can be inserted into a cell by:
• Microinjection
• Gene gun
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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