Molecular Biotechnology: Principles and Practices by Channarayappa (CRC) Providing a strong base in this emerging and highly promising field, Molecular Biotechnology: Principles and Practice strikes a balance between two important aspects of the science - the theory of molecular biology and the experimental approach to the study of biological processes. The main feature of this book is that it covers a wide range of molecular techniques in biotechnology and is designed to be a student- and teacher-friendly textbook. Each technique is described conceptually, followed by a detailed experimental account of the steps involved. The book can also serve as reference to the interested reader who is venturing into the field of biotechnology for the first time.
In the recent past, biotechnology has emerged as an important tool especially in the economic sectors of agriculture, livestock management, human health care and pharmaceutical industries, and for solving environmental and societal issues. As a result, many universities now offer various programs in biotechnology and related fields.
Molecular Biotechnology: Principles and Practices is intended as a student and teacher-friendly textbook aimed at providing undergraduate and postgraduate students with a strong base in this emerging and highly promising interdisciplinary science. It strikes a balance between two important aspects of the science—the theory of molecular biology and the experimental approach to the study of biological processes. The main feature of this book is its coverage of a wide range of molecular techniques in biotechnology. Each technique is described conceptually, followed by a detailed experimental account of the steps involved.
The topics covered include:
Channarayappa received his B.Sc. (Ag.), M.Sc. (Ag.), and Ph.D. degrees from the University of Agricultural Sciences, Bangalore. He then went on to earn a second Ph.D., in Genetics and Developmental Biology, from West Virginia University, USA, in 1990. He worked as a Postdoctoral Fellow and Postdoctoral Research Associate at the University of Texas MD Anderson Cancer Center, Houston. He received the Jawaharlal Nehru National Award for Outstanding Postgraduate Agricultural Research (1993) for significant research contribution in genetics and plant breeding, awarded by the ICAR, New Delhi. He has been teaching both undergraduate and postgraduate students of molecular biology and biotechnology for more than 20 years. He is the author of many scientific papers resulting from research conducted in plant sciences, mutation research, and cancer biology.
Excerpt: Biotechnology is an important tool that can be applied to various economic sectors such as the production of food crops, livestock management, human health care, chemical industries, and environmental management. Many universities, understanding the importance of biotechnology research and the need for qualified manpower to exploit such technologies, have started undergraduate and master's degree programs. With the burgeoning number of such courses, experts have realized the urgent need for developing a suitable curriculum, possibly in the form of a model textbook aimed at providing undergraduate and postgraduate students with a strong base in this emerging and highly promising interdisciplinary area.
Molecular Biotechnology: Principles and Practices is designed to balance between two important aspects of the science. The first aspect is the principles of molecular biology, which constitutes the theoretical knowledge pertaining to molecular biotechnology. The second aspect is practices in molecular biology, or the experimental approach to the study of biological processes. This book can serve as a textbook for both undergraduate and postgraduate students of molecular biology and biotechnology. It can also be used as a laboratory reference book in most research laboratories. The salient feature of this book is that it covers a wide range of molecular techniques in biotechnology and provides a source of information to readers at all levels. Concise and straightforward explanations of both theory and techniques associated with molecular/recombinant technologies are very few in literature. Most research articles in the field discuss either theory or techniques individually, but rarely explain both together and adequately. This makes life difficult for the student, teacher or researcher, who is new to the subject. Although several books on biotechnology have been published in the last decade, most are either very shallow or cover few areas in depth. Rarely do they cover the broad spectrum of topics which would provide enough information for understanding the subject or provide simple protocols for execution of molecular biology experiments. Realizing this deficiency, I have made an attempt to explain the basic concepts in biotechnology and the detailed steps of some important experiments in relatively simple terms. In my opinion, this book can help the reader to easily understand the subject and also execute the experiments very efficiently.
The book is divided into nine sections, containing 42 chapters and an appendix. In recent years, both the amount of molecular biology knowledge and its rate of growth have exploded. It is impossible to keep pace with this development and include references to all the work exhaustively. Therefore, in this book, only a few representative methods, which can provide standard protocols and general information on those techniques, are presented under each section. Alternative protocols provided for each technique make it more suitable for different laboratory conditions and systems used. Sources where additional information can be obtained are sufficiently quoted at the end of each chapter. Each chapter is adequately illustrated with computer graphics in an attempt to convey the practical as well as theoretical aspects of the techniques. It should, therefore be well suited for both the lecture room and the laboratory bench. The illustrations are adequately labeled and explains step-by-step, the procedure to be followed at each level of the process.
The recent advances in science and technology have driven the biological sciences into a new era. All of this can probably be traced back to when Anton van Leeuwenhoek, a Dutch dry-goods dealer, ground the first microscope lens. Through his newly invented glass, he discovered a previously unseen cellular world. The second half of the 21st century was a truly exciting time for molecular biologists. Many inventions, including new methods for analyzing proteins, DNA and RNA, fueled an explosion of information and enabled scientists to understand cells and multicellular organisms at the molecular level. Now we have molecular blueprints (genomic sequences) for many organisms. The main objective of modern science, however, is to know the nature of genetic material and to find the answers to questions like: Which genes determine specific characters? How do they get switched on and off spatially and temporally? How do we correct genetic defects? How can we best manipulate genomes?
The field of biotechnology, which emerged as a new discipline, was a result of the fusion of biology and technology. Biology is the science of all living organisms or their components, whereas technology deals with the physical–chemical properties and techniques (Chapters 313) applied to the production of biological products/services. The emergence of biotechnology has been possible mainly due to the revolutionary discoveries made in these two areas. Biotechnology has been defined in many ways by many organizations. Biotechnology may be broadly defined as "the controlled use of selected/manipulated biological systems or processes for the production of abundant/novel products or services". Therefore, the area covered under biotechnology is very vast and the techniques involved are widely divergent. Biotechnology can be applied in areas as diverse as agriculture, animal husbandry, medicine, environment, industry, and biological conservation.
Biotechnology is multidisciplinary by its very nature and encompasses several disciplines of basic sciences (e.g., genetics, biochemistry, molecular biology, chemistry, microbiology, immunology, cell and tissue culture, and physiology), engineering (processing technology, biochemical engineering, electronics and physical sciences) and also other disciplines like sociology, economics, politics, law and ethics
Although the term biotechnology is a recent development, its origin can be traced back to prehistoric times. Humans have been altering the genetic composition of plants for millennia –retaining seeds from the best crops and planting them in the following years, breeding and cross- breeding varieties to make them taste sweeter, grow bigger, last longer, etc. In this way, early agriculturists transformed the wild tomato (Lycopersicon), from a fruit the size of a peanut to today's giant, juicy and fleshy tomato. From a weedy plant called teosinte with an "ear" barely an inch long has emerged our foot-long ears of sweet, nutrient-rich, yellow corn. Man has also selected hundreds and thousands of new crop varieties by selection and hybridization. In ancient scripts, it has been documented that humans employed microorganisms as early as 5000 BC for making wine, vinegar, yogurt, leavened bread, etc. The discovery that fermentationconverted fruit juice into wine, milk into cheese and yogurt, and solutions of malt and hops into beer seems to have set in motion the study of biotechnology. The early animal breeders soon realized that different physical traits could be either magnified or lost by mating the appropriate pairs of animals, thereby engaging in the traditional manipulations of biotechnology. However, the use of microorganisms for the production of chemicals on a commercial scale begun during the First World War, and has recently been more fully exploited due to the advancement of modern biotechnology.
Modern biotechnology is innovative and quite different from the conventional practices. Traditional breeders made crosses only between related organisms whose genetic composition was compatible (genetically closer). Doing it this way involved the transfer of tens of thousands of genes (many genes were not required) after years of long selection procedure. By contrast, today's genetic engineers can transfer just a few desirable genes at a time, between species that are distantly related or not related at all. In other words, scientists can extract a desirable gene from virtually any living organism and insert it into virtually any other organism. They can put human gene into a plant or microorganism or in any other combination. For example, they can put a rat gene into lettuce to make a plant that produces vitamin C or insert a microbial toxin gene into cotton plants to make it insect-resistant. All this genetic manipulation became possible by the discovery of techniques of gene splicing and recombinant DNA technology. The engineered organisms which scientists produce by transforming genes between species are called "transgenic" organisms. Transgenic animals and several dozen transgenic food crops are currently in the market. Most of these crops are engineered to help farmers deal with age-old agriculture problems: good seeds, insects, diseases, nutrient composition, stress tolerance, etc.
The beginning of modern biotechnology can be traced back to 1865, when Gregor Mendel published the results from his experiments conducted on the garden pea on the inheritance of seven different physical traits. This and many other studies eventually led to the concept of the gene as the basic unit of heredity. Over the next century, many other researchers with sophisticated techniques and instruments contributed to the growth of modern biotechnology:
Before 6000 BCE Yeasts used to make wine and beer
About 4000 BCE Yeasts were used for making leavened bread
1866 Mendel published his research findings, experiments conducted on the garden pea, which led to the concept of the gene as the basic unit of heredity.
1869 Friedrich Miescher isolated nuclein, later shown to be DNA, from the nuclei of white blood cells.
1885 E. coli bacterial cells are identified and grown under controlled conditions.
1902 Archibald Garrod's report that the human disease "Alkaptonuria" behaves as a Mendelian recessive trait led to the suggestion that enzymes are encoded by genes.
1910 Thomas Hunt Morgan showed the first evidence of the presence of genes in chromosomes. He used microorganisms to treat sewage.
1912-14 Large-scale production of acetone, butanol and glycerol using bacteria.
1917 Karl Ereky coined the term "biotechnology".
1940 George Beadle and Edward Tatum hypothesized the concept of "one-gene-one-enzyme".
1943 Penicillin was produced on an industrial scale.
1944 Avery, Macleod and McCarty demonstrated that DNA, not protein, carries hereditary information. Penicillin was produced on a large scale for the first time.
1952 Alfred Hershey and Martha Chase demonstrated that the genes of a bacteriophage are made of DNA and are capable of directing the synthesis of new bacteriophage proteins.
1953 Watson and Crick determined the structure of the DNA double helix. They drew from the work of other scientists to propose that the molecule is an alpha double helix structure in which the two strands are both complementary and antiparallel to one another.
1955-65 The role of tRNA, mRNA, and rRNA as well as of DNA and RNA polymerases in gene function was elucidated.
1957 The Central Dogma, which states that hereditary information flows from DNA to RNA to protein, was put forth by Francis Crick and George Gamov.
1961 Marshall Nirenberg and Har Gobind Khorana correctly translated the genetic code.
1962 Uranium was mined with the aid of microbes (Canada).
1967 DNA ligase was isolated and identified.
1970 Stewart Lin and Werner Arber identified the first restriction endonucleases. In the same year researchers discovered the enzyme reverse transcriptase, which catalyzes the reaction in which DNA is transcribed from an RNA template.
1972 Khorana and co-workers synthesized an entire tRNA gene. Paul Berg created the first recombinant DNA molecule.
1973 Stanley Cohen, Herbert Boyer and colleagues constructed a functioning plasmid, containing genes, which confer resistance to both tetracycline and streptomycin; i.e., the establishment of recombinant DNA technology.
1976 Techniques were developed to determine the sequence of DNA.
1977 Somatostatin became the first human hormone to be synthesized by a bacterial cell as a result of transformation with human DNA.
1981 The use of monoclonal antibodies for diagnosis was approved in the USA.
1983 Approval was granted for the use of insulin produced by genetically-engineered microbes (GEMs).
1984 Animal interferons, produced by GEMs, were approved of for the protection of cattle against diseases.
1988 The polymerase chain reaction (PCR) method was published.
1990 Approval was granted in the USA for a trial of human somatic cell gene therapy.