Genetic Engineering

This book has a distinguishing feature of having condensed material with adequate information on genetic engineering especially of the microbes. The book covers almost all the topics of genetic engineering for the graduate, postgraduate students and young research scholars of biological sciences. The book is written as per syllabus of genetic engineering paper for Masters course in biotechnology, biochemistry, life sciences of most of the universities. The book is much useful for the students of Masters degree. Emphasis is given on the basic fundamentals. The book contains twelve chapters starting from ' Isolation, purification and estimation of nucleic acids' as chapter 1. The chapter describes general techniques for the isolation and purification of DNA as well as RNA. It also describes methods for quantitative estimation of the nucleic acids. The second chapter describes general characteristics of the vectors used in genetic engineering and also the general account of commonly used individual vectors. The chapter also describes expression vectors. The third chapter describes various commonly used restriction endonucleases. viz. Reverse transcriptase, DNA polymerase I, polynucleotide kinase, teminal dcoxynucleotidyl transferase, alkaline phosphatase, SI nuclease, DNA ligase etc. The fifth chapter describes electrophoresis for the separation of nucleic acids fragments. The sixth chapter is of cloning strategies. It describes construction of genomic DNA library, chromosomal walking, cDNA library, cDNA cloning. The seventh chapter describes DNA sequencing techniques and includes chemical modification method of Maxam and Gilbert, dideoxy sequencing method of Sanger, modifications of chain terminator sequencing, analysis of the sequencing data. The eighth chapter includes various methods of site directed mutagenesis. The ninth chapter describes polymerase chain reaction (PCR). It also includes primer designing and various types of polymerase chain reactions viz. reverse transcriptase polymerase chain reaction (RT-PCR), nested PCR, multiplex PCR etc. Besides, there are chapters 10, 11 and 12 on gene therapy, human genome and proteomics. At the end, glossary has been put which explains main terms used in genetic engineering. given in the beginning of each chapter that provides, at a glance, the contents of the whole chapter which offers a better learning mechanism. Each chapter is also presented with an introduction that covers the concept of the whole chapter in brief and offers clear understanding of the subject matter to the students. The author on the basis of his experience in teaching genetic engineering at the university level for more than a decade has offered the text in an easily understandable form to the postgraduate students. The book should be of invaluable help to the students, researchers and all those interested in understanding genetic engineering.