| Src was first identified in its viral form(v-Src) as a protein encoded by chicken Rous Sarcoma virus. The cellular homologue of v-Src is c-Src, and its protein product c-Src is located in the cytoplasm with 60KD. The members of Src family are non-receptor tyrosine kinases. Their make an important function in a complex network of intracellular signals, such as cell growth, proliferation, transformation, apoptosis, and cell adhesion, migration.The fact that Src was discovered as an oncogene suggests that Src could closely involvement in cancer. Many reports indicate that Src activity or Src transcriptional activation were elevated in human cancer. When Knock-out Src or added inhibitions, the malignant hyperplasia is reduced. And these suggest that the Src inhibitors could be used as anti-cancer agents. Furthermore, the Src family kinases are highly correlated with osteoporosis and brain development.The oncoprotein v-Src is similar to c-Src, and most functions of them is sameness, while some difference is important. V-Src can infection mammalian cell and leads to neoplastic transformation of cell, however, c-Src couldn't. In vitro, v-Src is activity and c-Src is inactive. Now, the researches are on Src activity in vivo. Thus these are not convenience for studies on the structure and chemical and Physical characteristics of Src. So through gene engineering technology, we clone v-Src gene into pGEX-KT, expressed in E.coli TGI and DH5 a after transformation and IPTG induction. The constructed plasmid containing GST-Src fused gene gave a high level of expression of the fused protein. Western blot analysis indicated that the fusion protein could specifically bind to anti-GST antibody. The SDS-PAGE analysis showed that the weight of the fusion protein is correct.With the v-Src protein, it will be studied on the structure of Src, the chemical and physical characteristics of Src, and Src activity in vitro. Furthermore, the studies may be on selection of anti-tumor agents, the antibody of Src. |
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