Textbook on Cloning, Expression and Purification of Recombinant Proteins

Module 1_Introduction.- Chapter 1. History of recombinant DNA technology.- Chapter 2. Overview of technology.- Chapter 3.          It's impact on research.- Module 2_Cloning basics.- Chapter 3. c-DNA library.- Chapter 4. PCR amplification.- Chapter 5. Restriction Digestion.- Chapter 6. Ligation.- Chapter 7. Colony screening.- Chapter 8. Transformation.- Chapter 9. Trouble-shooting.- Chapter 10. Problems.- Module 3_Choice of vectors (expression and cloning).- Chapter 11. Different types of vectors.- Chapter 12. Promoters.- Chapter 13. Antibiotic selection.- Chapter 14. Sequence confirmation.- Chapter 15. Trouble-shooting.- Chapter 16. Problems.- Module 4_Transformation and protein expression.- Chapter 17. Competent cell preparation.- Chapter 18. Transformation methods.- Chapter 19. Different bacterial strains (E. coli, Pseudomonas, Streptomyces).- Chapter 20. Optimizing protein expression (different inducers, temperature, concentration of inducers).- Chapter 21. Expression in insect cells.- Chapter 22. Trouble-shooting.- Chapter 23. Problem. Module 5_Protein purification.- Chapter  24. Lysis and extraction.- Chapter 25. Checking solubility and designing purification strategies.- Chapter 26. Databases and tools to determine physico-chemical properties of protein.- Chapter 27. Salting out and salting in methods.- Chapter 28. Dealing with proteins in inclusion bodies.- Chapter 29. Choice of buffer and dialysis.- Chapter 30. Overview of chromatography.- Chapter 31. Choice of chromatographic techniques with specific examples.- Chapter 32. Trouble-shooting.- Chapter 33. Problems.- Module 6_Affinity chromatography.- Chapter  34. Overview.- Chapter 35. His6.- Chapter 36. Cobalt.- Chapter 37. Streptavidin.- Chapter 38. MBP.- Chapter 39. GST.- Chapter 40. Trouble-shooting.- Chapter 41. Problems.- Module 7_Ion exchange chromatography.- Chapter 42. Overview.- Chapter 43. Cation

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.