基因,蛋白,免疫 课程之蛋白组学 6.doc

This lecture covers the production and use of transgenic animals A transgenic animal is one that has had a foreign gene inserted deliberately into its genome. They allow us to study gene function in the whole animal rather than in isolated cells or cell extracts They are widely used to create animal models of disease. This permits study of the evolution of the disease and the underlying mechanisms involved in the disease and to test the possible benefits or toxicity of novel therapies Transgenic animals have potential as sources of cells or tissue for transplant medicine They provide a means of producing recombinant proteins Learning objectives: By the end of your revision, you should understand what transgenic animals are and how they are produced. You should understand why, and be able to give examples of how, these are useful for research, have potential in medicine and have been used for the production of recombinant proteins? Background reading: Recombinant DNA JD Watson, M Gilman, J Witkowski, M Zoller Chapters 14 and 24 Human Molecular Genetics T Strachan, AP Read Chapters 5 and 20 Insights into cancer from transgenic mouse models MacLeod KE, Jacks T, J. Pathol. (1999) 187: 43-60. Genetic engineering of the donor as an approach to clinical xenotransplantation Cozzi E, Soin B, Holmes B, White D, Transplantation proceedings 32:2701-2703. Production of therapeutic proteins in the milk of transgenic livestock Colman A, Biochem, Soc. Symp. 63:141-147. Main steps involved Construction of expression vector cDNA of test protein vector: DNA for incorporation expression in host cells Production of suitable host cells Fertilised eggs Early embryos Totipotent or pluripotent Introduction of foreign gene into host cells Injection Retroviral infection Transfection Electroporation ± Selection Propagation in host animal ± Injection Gestation recipient female Selection of transgenic progeny Presence of transgene ± selection in progeny Analysis of trans