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目的:构建抗骨形成蛋白单链抗体并获得表达。方法:应用一段人工合成的含15个氨基酸的连接肽,采取亚克隆技术,将一株鼠抗BMP单克隆抗体的重链可变区(VH)的N端和轻链可变区(VL)的C端连接起来;将连接产物克隆入融合蛋白表达载体pEGX-4T-l,在大肠杆菌JM109中进行表达。结果:构建的单链抗体(ScFv)全长705bp,并获得融合表达产物约52×103u。结论:亚克隆方法是一种构建单链抗体快速、简便、可靠的方法。
Objective: To construct anti-osteogenic protein single-chain antibody and obtain its expression. Methods: The N-terminal and light chain variable region (VL) of the heavy chain variable region (VH) of a murine anti-BMP monoclonal antibody was subcloned by using a synthetic peptide containing 15 amino acids. The Ligation product was cloned into the fusion protein expression vector pEGX-4T-1 and expressed in E. coli JM109. Results: The full-length single-chain antibody (ScFv) was 705bp and the fusion protein was about 52 × 103u. Conclusion: The subcloning method is a rapid, simple and reliable method to construct single chain antibody.