Chapter 9

whipmellificiumBiotechnology

Feb 20, 2013 (4 years and 5 months ago)

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Medical Technology Department, Faculty of Science, Islamic University
-
Gaza

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

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Figure 9.1.1

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Figure 9.1.2

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Table 9.1.1

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Table 9.1.2

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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

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Cut specific sequences of DNA


Destroy bacteriophage DNA in bacterial cells


Cannot digest (host) DNA with methylated cytosines

Restriction Enzymes

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Figure 9.2

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Carry new DNA to desired cell


Shuttle vectors can exist in several different species


Plasmids and viruses can be used as vectors

Vectors

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Vectors

Figure 9.3

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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)

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PCR

Figure 9.4.1

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PCR

Figure 9.4.2

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Transformation


Electroporation


Protoplast fusion

DNA can be inserted into a cell by:

Figure 9.5b

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Microinjection


Gene gun

DNA can be inserted into a cell by:

Figure 9.6 & 7

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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

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Genetic Engineering

Figure 9.11.1

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Genetic Engineering

Figure 9.11.2

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Making a Gene Product

Figure 9.12.1

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Making a Gene Product

Figure 9.12.2

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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

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Making a Gene Product

Figure 9.13

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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

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Random Shotgun Sequencing

Figure 9.14

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Understanding of
DNA


Sequencing
organisms'
genomes


DNA fingerprinting
for identification

Scientific Applications

Figure 9.16

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Southern Blotting

Figure 9.15.1

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Southern Blotting

Figure 9.15.2

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Southern Blotting

Figure 9.15.3

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Agricultural Applications

Table 9.2

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Genetic Engineering Using
Agrobacterium

Figure 9.18

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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