1st Edition

Techniques in Genetic Engineering




ISBN 9781482260892
Published May 8, 2015 by CRC Press
346 Pages 104 B/W Illustrations

USD $155.00

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Book Description

Although designed for undergraduates with an interest in molecular biology, biotechnology, and bioengineering, this book—Techniques in Genetic Engineering—IS NOT: a laboratory manual; nor is it a textbook on molecular biology or biochemistry. There is some basic information in the appendices about core concepts such as DNA, RNA, protein, genes, and genomes; however, in general it is assumed that the reader has a background on these key issues.

Techniques in Genetic Engineering briefly introduces some common genetic engineering techniques and focuses on how to approach different real-life problems using a combination of these key issues. Although not an exhaustive review of these techniques, basic information includes core concepts such as DNA, RNA, protein, genes, and genomes. It is assumed that the reader has background on these key issues. The book provides sufficient background and future perspectives for the readers to develop their own experimental strategies and innovations.

This easy-to-follow book presents not only the theoretical background of molecular techniques, but also provides case study examples, with some sample solutions. The book covers basic molecular cloning procedures; genetic modification of cells, including stem cells; as well as multicellular organisms, using problem-based case study examples.

Table of Contents

Introduction to Genetic Engineering
Tools of Genetic Engineering

Restriction Endonucleases
Vectors
Modifying Enzymes
Basic Principles of Cloning
Problem Session
DNA Libraries
Introduction
Genomic DNA Libraries
cDNA Libraries
Library Screening
Monitoring Transcription
Problem Session
Protein Production and Purification
Expression Vectors and Recombinant Protein Expressions
In Vitro Transcription and Translation
Bacterial Expression of Proteins
Expressions in Yeast
Expressions in Insect Cells
Expressions in Plant Cells
Expressions in Mammalian Cells
Purification of Proteins
Post-Translational Modifications of Proteins
Problem Session
Mutagenesis
Mutagenesis
Deletion Studies
Site-Directed Mutagenesis
Random Mutagenesis
Directed Evolution, Protein Engineering, and Enzyme Engineering
Problem Session
Protein-Protein Interactions
Introduction
GST Pull-Down-Based Interaction Assay
Co-Immunoprecipitation
The Yeast Two-Hybrid Assay
Fluorescence Resonance Energy Transfer
Problem Session
Cell Culture
Introduction
Genetic Manipulation of Cells
Reporter Genes
Types of Transfection
Level of Expression
Problem Session
Genetic Manipulation of Stem Cells and Animals
Stem Cell Technology and Knockout Cells
Transgenic Animals
RNA Interference and MicroRNAs
Animal Cloning
Pharm Animals
Gene Therapy
Genome Editing
Problem Session
Genetic Manipulation of Plants
Monocotyledons, Dicotyledons, and Commercial Crops
Plant Manipulation Methods
Future Trends in Transgenic Plants
Problem Session
Today and the Future
Bioinformatics and the Omics Age
Synthetic Biology and Unnatural Amino Acids
Optogenetics
What Is Next?
Problem Session
References
Glossary
A: DNA Techniques
A.1 DNA Gel Electrophoresis
A.2 Nucleic Acid Blotting
A.3 Polymerase Chain Reaction
A.4 Real-Time PCR
A.5 DNA Sequencing
A.6 Next Generation Sequencing
B: Protein Techniques
B.1 SDS-PAGE
B.2 Western Blotting
B.3 2D Gel Electrophoresis and Proteomics
B.4 Immunofluorescence and Immunohistochemistry
Supplement Information for End-of-Chapter Questions
1 Nucleotides and Nucleic Acids
2 Genetic Coding Tables
3 Amino Acids
4 Calculations Regarding Nucleic Acids, Amino Acids, and Proteins
5 Compatible Overhangs
6 Genotypes of Frequently Used Laboratory Strains of Bacteria and Yeast
7 Methylation Sensitivities of Common Restriction Enzymes
8 Primary and Secondary Antibody Examples
9 Fluorophores
10 Radioisotopes in Biology
Answers to End-of-Chapter Questions
1 Chapter 2 Answers
2 Chapter 3 Answers
3 Chapter 4 Answers
4 Chapter 5 Answers
5 Chapter 6 Answers
6 Chapter 8 Answers
7 Chapter 9 Answers

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Author(s)

Biography

Prof. Işıl AKSAN KURNAZ has received her BSc from Bogazici University, Department of Molecular Biology and Genetics, after which she did her PhD with Dr. Colin Goding in Marie Curie Research Institute, UK (closed down in 2010) and University of London, Institute of Cancer Research. Following her degree, she has worked as a lecturer in Bogazici University (1999-2000), and a postdoctoral researcher with Prof. Andrew D. Sharrocks in University of Manchester (2000-2002). She was one of the two young assistant professors to be then recruited by Yeditepe University in Istanbul, Turkey, so as to set up biotechnology and genetics laboratories as well as the Department of Genetics and Bioengineering in 2003. She continued to work there as a professor and principle investigator of the Molecular Neurobiology Laboratory (aka AxanLab, https://www.facebook.com/AxanLab) until 2014, when she relocated her laboratory to Gebze Institute of Technology (GYTE) in Kocaeli, Turkey.

Prof. Kurnaz is the recipient of the L’Oreal Turkey Young Female Investigator Award (as a local counterpart of the international For Women in Science programme) in 2006, and the GEBIP Award (Genc Bilim Insanlarini Destekleme Programi / Distinguished Young Investigator Award) of the Turkish Academy of Sciences (TUBA) in 2007. She has also participated in the European Young Academy brainstorming meeting organized by ALLEA and ESF in Vienna, 2009. She has been involved in several international grants such as COST and FP7 Regpot projects, as well as many national grants from TUBITAK, Turkish Fight with Cancer Foundation, Brain Research Society – Lundbeck, Novartis among many others. She has published numerous papers, to which she has received over 500 citations, with a current h index of 9.