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Genetic Engineering – Basics, New Applications and Responsibilities GENETIC ENGINEERING – BASICS, NEW APPLICATIONS AND RESPONSIBILITIES Edited by Hugo A. Barrera-Saldaña Genetic Engineering – Basics, New Applications and Responsibilities Edited by Hugo A. Barrera-Saldaña Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2011 InTech All chapters are Open Access distributed under the Creative Commons Attribution 3.0 license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. As for readers, this license allows users to download, copy and build upon published chapters even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. Publishing Process Manager Gorana Scerbe Technical Editor Teodora Smiljanic Cover Designer InTech Design Team Image Copyright Aspect3D, 2011. Used under license from Shutterstock.com First published January, 2011 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from [email protected] Genetic Engineering – Basics, New Applications and Responsibilities, Edited by Hugo A. Barrera-Saldaña p. cm. ISBN 978-953-307-790-1 Contents Preface IX Part 1 Technology 1 Chapter 1 Expression of Non-Native Genes in a Surrogate Host Organism 3 Dan Close, Tingting Xu, Abby Smartt, Sarah Price, Steven Ripp and Gary Sayler Chapter 2 Gateway Vectors for Plant Genetic Engineering: Overview of Plant Vectors, Application for Bimolecular Fluorescence Complementation (BiFC) and Multigene Construction 35 Yuji Tanaka, Tetsuya Kimura, Kazumi Hikino, Shino Goto, Mikio Nishimura, Shoji Mano and Tsuyoshi Nakagawa Part 2 Application 59 Chapter 3 Thermostabilization of Firefly Luciferases Using Genetic Engineering 61 Natalia Ugarova and Mikhail Koksharov Chapter 4 Genetic Engineering of Phenylpropanoid Pathway in Leucaena leucocephala 93 Bashir M. Khan, Shuban K. Rawal, Manish Arha, Sushim K. Gupta, Sameer Srivastava, Noor M. Shaik, Arun K. Yadav, Pallavi S. Kulkarni, O. U. Abhilash, SantoshKumar, Sumita Omer, Rishi K. Vishwakarma, Somesh Singh, R. J. Santosh Kumar, Prashant Sonawane, Parth Patel, C. Kannan, Shakeel Abbassi Chapter 5 Genetic Engineering of Plants for Resistance to Viruses 121 Richard Mundembe, Richard F. Allison and Idah Sithole-Niang Chapter 6 Strategies for Improvement of Soybean Regeneration via Somatic Embryogenesis and Genetic Transformation 145 Beatriz Wiebke-Strohm, Milena Shenkel Homrich, Ricardo Luís Mayer Weber, Annette Droste and Maria Helena Bodanese-Zanettini VI Contents Chapter 7 Genetic Engineering and Biotechnology of Growth Hormones 173 Jorge Angel Ascacio-Martínez and Hugo Alberto Barrera-Saldaña Part 3 Biosafety 197 Chapter 8 Genetically Engineered Virus-Vectored Vaccines – Environmental Risk Assessment and Management Challenges 199 Anne Ingeborg Myhr and Terje Traavik Part 4 Responsibility 225 Chapter 9 Genetic Engineering and Moral Responsibility 227 Bruce Small Preface In the last three decades since the application of genetics to plants and animals, we have witnessed impressive advances best illustrated by the fact that almost one-tenth of all cultivated land on our planet is now planted with transgenic crops. Also, although no transgenic animals can be found in the prairies, some do live on specialized farms, replacing bioreactors from biotech facilities in the production of therapeutic proteins. While the targets of the first efforts of genetic engineering were to increase plant resistance to pests and herbicides, some ingenious and provocative applications also started emerging, such as longer lasting fruit on the shelf and mice, even pet animals, expressing the sea medusa green fluorescent protein.Genetic engineering has proven that it is not a threat to mankind but rather a powerful tool for solving not only food shortages, especially by reducing losses due to pests and by contributing to the development of inexpensive and safer fertilizers, but also for decreasing the shortage of sophisticated biologicals from natural sources and for coping with the explosive demand of these in medicine. A good example are antigens and therapeutics, which are now produced even by cows in modern biotech farms. At the same time, we are exposed to novel applications of genetic engineering in practically all fields. This book illustrates some of these applications, such as thermo- stabilization of luciferase; engineering of the phenylpropanoid pathway in a species of high demand for the paper industry; more efficient regeneration of transgenic soybean; viral resistant plants; and a novel approach for rapidly screening, in the test tube, properties of newly discovered animal growth hormones. To make the technology more user-friendly and easy to understand, two chapters focus on the basics of making the expression of transgenes in plants and biotech hosts possible. They also illustrate the state-of-the-art tools (mainly expression vectors) that are capable of coping with the hosts´ requirements for expressing their own genes. Finally, there are chapters concerned with safety issues in manipulating plants, viruses, and introducing genetically modified organisms into the environment, and with how to raise consciousness of the great responsibility we now carry to use genetic engineering wisely and planet-friendly. X Preface The book contributes chapters on the basics of genetic engineering, on applications of the technology to attempt to solve problems of greater importance to both society and industry, and comes to a close by reminding us of the moral responsibility we have to always keep in mind, that nature is a very fragile equilibrium and that we have already put it at risk. We should always pay attention to the ethical, moral and environmental consequences of applications that have not been tested enough in the laboratory and in controlled field facilities to avoid unexpected and unintentional harm to our and other species and as well as the environment. Prof. Dr. Hugo A. Barrera-Saldaña Professor, Department of Biochemistry and Molecular Medicine UANL School of Medicine Monterrey, Director, Vitaxentrum Monterrey, México Part 1 Technology 1 Expression of Non-Native Genes in a Surrogate Host Organism Dan Close, Tingting Xu, Abby Smartt, Sarah Price, Steven Ripp and Gary Sayler Center for Environmental Biotechnology, The University of Tennessee, Knoxville USA 1. Introduction Genetic engineering can be utilized to improve the function of various metabolic and functional processes within an organism of interest. However, it is often the case that one wishes to endow a specific host organism with additional functionality and/or new phenotypic characteristics. Under these circumstances, the principles of genetic engineering can be utilized to express non-native genes within the host organism, leading to the expression of previously unavailable protein products. While this process has been extremely valuable for the development of basic scientific research and biotechnology over the past 50 years, it has become clear during this time that there are a multitude of factors that must be considered to properly express exogenous genetic constructs. The major factors to be considered are primarily due to the differences in how disparate organisms have evolved to replicate, repair, and express their native genetic constructs with a high level of efficiency. As a result, the proper expression of exogenous genes in a surrogate host must be considered in light of the ability of the replication and expression machinery to recognize and interact with the gene of interest. In this chapter, primary attention will be given to the differences in gene expression machinery and strategies between prokaryotic and eukaryotic organisms. Factors such as the presence or absence of exons, the functionality of polycistronic expression systems, and differences in ribosomal interaction with the gene sequence will be considered to explain how these discrepancies can be overcome when expressing a prokaryotic gene in a eukaryotic organism, or vice versa. There are, of course, additional concerns that are applicable regardless of how closely related the surrogate host is to the native organism. To properly prepare investigators for the expression of genes in a wide variety of non-native organisms, concerns such as differences in the codon usage bias of the surrogate versus the native host, as well as how discrepancies in the overall GC content of each organism can affect the efficiency of gene expression and long term maintenance of the construct will be considered in light of the mechanisms employed by the
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