Gene cloning and manipulation
著者
書誌事項
Gene cloning and manipulation
Cambridge University Press, c1995
- hbk
- pbk.
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注記
Includes bibliographical references (p. 195-200) and index
内容説明・目次
内容説明
This text gives a broad, but concise, coverage of gene cloning and manipulation, suitable for undergraduates and beginning graduate students. Assuming only general biochemical knowledge, it stresses the concepts underlying particular types of cloning vector, and uses examples to illustrate them, rather than simply presenting a mass of detailed lists and vector maps. The book starts by describing the principles behind cloning DNA in E. coli, the enzymes used, the range of cloning vectors available, and how to screen libraries to find particular clones. The author shows how PCR can be used as an alternative, or complementary, approach. He then goes on to describe how sequences can be exploited, after cloning and identification, by site-directed mutagenesis and over-expression. The book finishes with a detailed presentation of the genetic manipulation of other organisms, including other bacteria, yeast, plants, insects, and mammals.
目次
- Part I. The Tools for the Job: 1. Introduction
- 2. Cutting
- 3. Modification
- 4. Ligation
- 5. Purification of plasmid DNA from E. Coli
- 6. Gel electrophoresis of nucleic acids
- 7. Oligonucleotide synthesis
- Part II. Simple Cloning: 8. The basic experiment
- 9. Vectors
- transformation and hosts
- 10. Modifications
- 11. Linkers, adaptors and cassettes
- Part III. Other vector systems for E. Coli
- 12. Introduction
- 13. Bacteriophage M13 and its uses
- 14. Bacteriophage lambda
- 15. Cosmids
- 16. Bacteriophage P1
- Part IV. Making Libraries: 17. Introduction
- 18. Genomic libraries
- 19. cDNA libraries
- 20. Specialized libraries
- Part V. Screening Libraries: 21. Introduction
- 22. Screening for coding function
- 23. Screening for other functions-reporter genes
- Part VI. Polymerase Chain Reaction: 24. The basic technique
- 25. Modifications
- 26. Precautions and drawbacks
- 27. Modification and mutagenesis
- 28. Introduction
- 29. Alteration of restriction sites
- 30. Insertions and deletions
- 31. Point mutations - early methods
- 32. Oligonucleotide mutagenesis
- 33. Choosing the right mutations
- 34. Inactivating genes
- Part VII. Use of Cloned DNA: 35. As DNA
- 36. Synthesis of RNA
- 37. Synthesis of protein
- Part VIII. Using Other Organisms: 38. Gram-negative bacteria
- 39. Gram-positive bacteria
- 40. Fungi
- 41. Chlamydomonas
- 42. Vascular plants
- 43. Organelle transformation
- 44. Insects
- 45. Mammals
- References and further reading
- Index.
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