Textbook on Cloning, Expression and Purification of Recombinant Proteins

This book is immensely useful for graduate students as well as researchers to understand the basics of molecular biology and Recombinant DNA Technology. It provides a comprehensive overview of different approaches for the synthesis of recombinant proteins from E. coli including their cloning, expression and purification. Recent advances in genomics, proteomics, and bioinformatics have facilitated the use of Recombinant DNA Technology for evaluating the biophysical and biochemical properties of various proteins. The book starts with an introductory chapter on gene cloning, protein expression and purification and its implication in current research and commercial applications. Each chapter provides a lucid set of principles, tools and techniques for both students and instructors. The protocols described have been aptly exemplified, and troubleshooting techniques have been included to aid better understanding. Moreover, the set of questions at the end of each chapter have been particularly formulated to help effective learning.

Provides overview of recombinant DNA technology, including gene cloning, protein expression and purification In depth description of various chromatographic techniques that are utilized to purify recombinant proteins Discusses techniques that are employed to study the biophysical properties of various proteins

Auteur
Dr. Kakoli Bose is currently working as Scientific Officer 'G' at Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Mumbai, India. She is known for her expertise in a diverse range of fields from Protein Biochemistry, Enzymology, Structural Biology, Biophysics to Cell Biology and Computational Biology. Her research activities are focused on elucidation of the role of macromolecules that are involved in pathways of apoptosis and their associated diseases. She is an active member of many elite scientific societies and associations, including Biochemical Society, UK, Indian Biophysical Society, Indian Association for Cancer Research and American Chemical Society. Dr. Bose is also a core member of the Editorial Board of coveted international journals such as Bioscience Reports and the Biochemical Journal, Portland Press, UK. She is a recipient of many prestigious awards, including the National Women Bioscientist Award from Govt. of India. She has a teaching experience of more than 10 years in the fields of Biochemistry, Biophysics, and Structural Biology. She has several publications in reputed international journals to her credit. Furthermore, her publication involving a four-state equilibrium unfolding process remains a landmark and pioneering work till date.

Contenu

A Brief Introduction to Recombinant DNA Technology.- Cloning and Gene Manipulation.- Selection of Cloning and Expression Plasmid Vectors.- Transformation and Protein Expression.- Introduction to Recombinant Protein Purification.- Protein Purification by Affinity Chromatography.- Protein Purification by Ion Exchange Chromatography.- Gel Filtration Chromatography.- Protein Purification by Reversed Phase Chromatography and Hydrophobic Interaction Chromatography.- Purification of Difficult Proteins.- Protein Quantitation and Detection.