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GENETIC MANIPULATION OF DNA AND PROTEIN – EXAMPLES FROM CURRENT RESEARCH

GENETIC MANIPULATION OF DNA

Edited by David Figurski .

462 pages . 
Open Access .


This diverse collection of research articles is united by the enormous power of modern molecular genetics. The current period is an exciting time both for researchers and the  curious who want to know more about genetic approaches to solving problems.
This volume is noteworthy. Every author accomplished two important objectives: (1)  making the field and the particular research described accessible to a large audience  and (2) explaining fully the genetic tools and approaches that were used in the  research. One fact stands out – the importance of a genetic approach to addressing a  problem. I encourage you to read several chapters. You will feel the excitement of the  scientists, and you will learn about an area of research with which you may not be  familiar. Perhaps most importantly, you will understand the genetic approaches; and  you will appreciate their importance to the research.
Anyone can benefit from reading these chapters – even those of you who have a solid  foundation in modern molecular genetics. This is an eclectic mix of topics (only the  surface has been scratched). These chapters are valuable, not only because they reflect  the current state of the art and are easy to read, but also because they are concise  reviews. The variety will provide you with new knowledge to be sure, but it may also  affect your own thoughts about a problem. Thinking about a topic very different from  the one you are considering can stimulate fresh and often unconventional ideas.
We all know that the code for all life on the planet is in DNA and RNA. The purpose  of genetics is to decipher life’s information – to understand why the genome codes for  its various functions. Much of the work in this volume is geared to manipulating DNA  with that knowledge, not only to provide clues about a function, but also to test an  idea or to change a protein to learn how it works or to make it work better.
For a time, the field of molecular genetics was concerned with a few manipulable  model organisms. This was necessary to answer basic questions like “How does a gene  work?” Now modern molecular genetics has given us the confidence to explore the  unknowns in the diversity of life, including complex organisms, like humans. We may  need to adapt or develop genetic tools (see the contents section on tools). We have  already learned that many of the “paradigms” of the model organisms do not apply to  other organisms.
“Manipulate” is a problem word in genetics for some people. This volume has another  purpose - to be accessible to those who fear the power of genetics. Those of us who  know modern genetics understand that the current precision of genetically modified  food, for example, is far safer than the unknowns of genetic crosses, a technology that  is strangely acceptable. We have ourselves to blame for the apparent mystery and the  public’s misperceptions. Too often we discuss our work with our colleagues but fail to  explain our work to the public.
By making these chapters freely available to everyone and by the authors clearly  describing the question being asked and the approach taken to answer it, this book is  partly addressing that concern. People who fear genetics should take comfort in the  dissemination of knowledge about this science. Scientists have the same concerns as  the public. The more who understand genetics, the more there will be vigilance.
This collection of research articles is testimony to the optimism in the field. Both major  and minor problems can be solved. For example, genetics will likely be a part of the  solution to hunger, and genetically engineered microorganisms may help solve the  problem of global warming. Basic research (see the contents sections on basic research  and the development of approaches and tools) is difficult to explain, but it is vitally  important for any progress. Genetics will help alleviate suffering by leading to new  therapies for disease (see the contents section on disease-related research), and it can  generate improved or new molecular activities (see the contents section on applied  research).
With a complete understanding of genetics, humankind will reach an important new  stage. Humans will be able to change their own genes. Of course, evolution will  continue to be an agent of genetic change; but it is slow in humans, and it acts on  populations. With the knowledge of genetics, humans will be able to direct change  (like the curing of a disease) to an individual; and it can be rapid.
You will be exposed to investigations on bacteria, archaea, fungi, mitochondria, and  higher eukaryotes, including humans. You will learn about various genetic  approaches, including specific alteration of amino acid residues in proteins, gene  fusions, cysteine- and alanine-scanning mutagenesis, recombineering, cloning by  “capturing” large segments of DNA, transposable elements, and allelic exchange. The  chapters are all very readable, and again I encourage you to sample more than one.

David Figurski
Professor of Microbiology & Immunology at
 Columbia University,
USA
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CONTENTS : 

Section 1 Molecular Genetics in Basic Research .


  1 Site-Directed Mutagenesis and Yeast Reverse 
2-Hybrid-Guided Selections to Investigate the Mechanism of Replication Termination 3 Deepak Bastia, S. Zzaman and Bidyut K. Mohanty

  2 Biochemical Analysis of Halophilic Dehydrogenases 
Altered by Site-Directed Mutagenesis 17 J. Esclapez, M. Camacho, C. Pire and M.J. Bonete

  3 Targeted Mutagenesis in the Study of the Tight Adherence 
(tad) Locus of Aggregatibacter actinomycetemcomitans 43 David H. Figurski, Daniel H. Fine, Brenda A. Perez Cheeks, Valerie W. Grosso, Karin E. Kram, Jianyuan Hua, Ke Xu and Jamila Hedhli
Chapter 4 Directed Mutagenesis of Nicotinic Receptors to Investigate Receptor Function 71
Jürgen Ludwig, Holger Rabe, Anja Höffle-Maas, Marek Samochocki, Alfred Maelicke and Titus Kaletta


  5 Site-Directed Mutagenesis as a Tool to Characterize 
Specificity in Thiol-Based Redox Interactions Between Proteins and Substrates 91 Luis Eduardo S. Netto and Marcos Antonio Oliveira

  6 Protein Engineering in Structure-Function Studies 
of Viper's Venom Secreted Phospholipases A2 107 Toni Petan, Petra Prijatelj Žnidaršič and Jože Pungerčar

  7 Site-Directed Mutagenesis in the Research of Protein Kinases 
- The Case of Protein Kinase CK2 133 Ewa Sajnaga, Ryszard Szyszka and Konrad Kubiński

Chapter 8 Directed Mutagenesis in Structure Activity Studies 
of Neurotransmitter Transporters 167 Jane E. Carland, Amelia R. Edington, Amanda J. Scopelliti, Renae M. Ryan and Robert J. Vandenberg

  9 Site-Directed Mutagenesis as a Tool for Unveiling 
Mechanisms of Bacterial Tellurite Resistance 185 José Manuel Pérez-Donoso and Claudio C. Vásquez

Section 2 Molecular Genetics in Disease-Related Research .


  10 A Mutagenesis Approach for the Study of 
the Structure-Function Relationship of Human Immunodeficiency Virus Type 1 (HIV-1) Vpr 203 Kevin Hadi, Oznur Tastan, Alagarsamy Srinivasan and Velpandi Ayyavoo

  11 New Insights into the Epithelial Sodium Channel 
Using Directed Mutagenesis 221 Ahmed Chraibi and Stéphane Renauld

  12 Use of Site-Directed Mutagenesis in the Diagnosis, 
Prognosis and Treatment of Galactosemia 233 M. Tang, K.J. Wierenga and K. Lai

Chapter 13 Inherited Connective Tissue Disorders of Collagens: 
Lessons from Targeted Mutagenesis 253 Christelle Bonod-Bidaud and Florence Ruggiero

Section 3 Molecular Genetics in Applied Research .


  14 Biological Activity of Insecticidal Toxins: Structural Basis, 
Site-Directed Mutagenesis and Perspectives 273 Silvio Alejandro López-Pazos and Jairo Cerón

Chapter 15 Site-Directed Mutagenesis as Applied to Biocatalysts 303 
Juanita Yazmin Damián-Almazo and Gloria Saab-Rincón

Section 4 New Tools or Approaches for Molecular Genetics .


  16 Using Cys-Scanning Analysis Data 
in the Study of Membrane Transport Proteins 333 Stathis Frillingos

  17 Site-Directed Mutagenesis Using 
Oligonucleotide-Based Recombineering 361 Roman G. Gerlach, Kathrin Blank and Thorsten Wille

  18 Studying Cell Signal Transduction 
with Biomimetic Point Mutations 381 Nathan A. Sieracki and Yulia A. Komarova

  19 Using Genetic Reporters to Assess Stability and Mutation 
of the Yeast Mitochondrial Genome 393 Shona A. Mookerjee and Elaine A. Sia

  20 Site-Directed and Random Insertional Mutagenesis 
in Medically Important Fungi 417 Joy Sturtevant

  21 Recombineering and Conjugation as Tools 
for Targeted Genomic Cloning 437 James W. Wilson, Clayton P. Santiago, Jacquelyn Serfecz and Laura N. Quick . 


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