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目的 :获得pS2融合蛋白。方法 :RT PCR法从乳腺癌组织中扩增pS2片段 ,定向克隆到PMS 31b载体上 ,重组质粒PMS pS2转化温度敏感型细菌POP2 136 ,使之高效表达融合蛋白MS2 pS2。结果 :免疫组织化学试验 (IHC)结果表明 :纯化的融合蛋白MS2 pS2能与抗pS2进口单抗特异反应。酶联免疫吸附试验 (ELISA)及Westernblotting试验表明 :MS2 pS2能与pS2C端 31个氨基酸合成肽抗血清特异反应。以此融合蛋白为免疫原得到的抗血清可用于免疫组织化学分析。结论 :pS2融合蛋白表达是正确的 ,亦为进一步研究pS2生物学功能及制备抗pS2单抗奠定了基础。
Purpose: To obtain pS2 fusion protein. METHODS: The pS2 fragment was amplified by RT-PCR from breast cancer tissues and cloned into PMS 31b vector. The recombinant plasmid pMS pS2 was transformed into temperature-sensitive bacteria POP2 136 to express the fusion protein MS2 pS2 efficiently. Results: Immunocytochemistry (IHC) results showed that the purified fusion protein MS2 pS2 specifically reacted with anti-pS2 mAb. Enzyme-linked immunosorbent assay (ELISA) and Western blotting showed that MS2 pS2 reacted specifically with the 31 amino acid synthetic peptide antisera of pS2C. The antiserum obtained by using the fusion protein as an immunogen can be used for immunohistochemical analysis. CONCLUSION: The expression of pS2 fusion protein is correct, which lays the foundation for further study on the biological function of pS2 and the preparation of anti-pS2 monoclonal antibody.