Chapter 9, part A

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20 Φεβ 2013 (πριν από 3 χρόνια και 10 μήνες)

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Microbiology

B.E Pruitt & Jane J. Stein

AN INTRODUCTION

EIGHTH EDITION

TORTORA
• FUNKE • CASE

Chapter 9, part A

Biotechnology and Recombinant
DNA

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

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