Techniques in Genetic Engineering: 1st Edition (Hardback) book cover

Techniques in Genetic Engineering

1st Edition

By Isil Aksan Kurnaz

CRC Press

346 pages | 104 B/W Illus.

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pub: 2015-05-08
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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

About the Author

Prof. Isil 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.

Subject Categories

BISAC Subject Codes/Headings:
MED009000
MEDICAL / Biotechnology
SCI029000
SCIENCE / Life Sciences / Genetics & Genomics
SCI043000
SCIENCE / Research & Methodology