Biomedical Engineering Reference
In-Depth Information
Cloning in
Bacteria
Other Than
E.coli
Chapter 8
Getting DNA into bacteria
Cloning in Gram-negative bacteria
Cloning in Gram-positive bacteria
Why clone in fungi
Vectors for use in fungi
Expression of cloned DNA
Two hybrid system
Analysis of the whole genome
Chapter 9
Cloning in
Yeast &
Other
Fungi
Transformation of animal cells
Use of non-replicating DNA
Replication vectors
Viral transduction
Gene
Transfer
To Animal
Cells
Chapter 10
Transgenic mice
Other transgenic mammals
Transgenic birds, fish,
Xenopus
Transgenic invertebrates
Chapter 11
Genetic
Manipulation
of Animals
Handling plant cells
Agrobacterium
-mediated transformation
Direct DNA transfer
Plant viruses as vectors
Chapter 12
Genetic
Manipulation
of Plants
Inducible expression systems
Site-specific recombination
Gene inhibition
Insertional mutagenesis
Gene tagging
Entrapment constructs
Chapter 13
Advanced
Techniques
for Gene
Manipulation
in Plant and
Animals
Fig. 1.3
'Roadmap' of the advanced
techniques in gene manipulation and
their application to organisms other
than
E. coli
.
In the second half of the topic the specialist tech-
niques for cloning in organisms other than
E. coli
are described (Fig. 1.3). Each of these chapters can
be read in isolation from the other chapters in this
section, provided that there is a thorough under-
standing of the material from the first half of the
topic. Chapter 8 details the methods for cloning in
other bacteria. Originally it was thought that some
of these bacteria, e.g.
B. subtilis
, would usurp the
position of
E. coli.
This has not happened and gene
manipulation techniques are used simply to better
understand the biology of these bacteria. Chapter 9
focuses on cloning in fungi, although the emphasis
is on the yeast
Saccharomyces cerevisiae.
Fungi are
eukaryotes and are useful model systems for invest-
igating topics such as meiosis and mitosis, control of
cell division, etc. Animal cells can be cultured like
microorganisms and the techniques for cloning in
them are described in Chapter 10. Chapters 11 and
12 are devoted to the intricacies of cloning in animal
and plant representatives of higher eukaryotes and
Chapter 13 covers some cutting-edge techniques for
these same systems.
The concluding chapter is a survey of the dif-
ferent applications of recombinant DNA techno-
logy that are being exploited by the biotechnology
industry. Rather than going through application
after application, we have opted to show the inter-
play of different technologies by focusing on six
themes:
• Nucleic acid sequences as diagnostic tools.
• New drugs and new therapies for genetic diseases.
• Combating infectious disease.
• Protein engineering.
• Metabolic engineering.
• Plant breeding in the twenty-first century.
By treating the topic in this way we have been able to
show the interplay between some of the basic tech-
niques and the sophisticated analysis now possible
with genome sequence information.
