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目的 选取调控补体活化中心环节C3转化酶的两种膜调节蛋白CD4 6和CD5 5 ,设计构建了新型的GPI锚固型CD4 6 /CD5 5嵌合分子。方法 以CD4 6cDNA和CD5 5cDNA为模板 ,通过PCR及PCR重叠延伸技术 (SOE)得到CD4 6 /CD5 5嵌合cDNA。为避免双分子连接后的空间结构和功能的变化 ,在CD4 6与CD5 5之间引入了多肽氨基酸接头 (Linker) ,然后克隆构建GPI锚固型CD4 6 /CD5 5 BluescriptM13克隆载体。结果 获得了阅读框完整、连接部位正确的GPI锚固型CD4 6 /CD5 5嵌合分子cDNA。结论 本研究为研制开发新一代的多功能、多靶点的补体抑制剂奠定了基础
Objective To design and construct a new type of GPI anchored CD4 6 / CD5 5 chimeric molecule by selecting two membrane regulatory proteins, CD4 6 and CD5 5, which regulate the C3 converting enzyme in the activation center of complement. Methods Using CD4 6 cDNA and CD5 5 cDNA as templates, CD4 6 / CD5 5 chimeric cDNA was obtained by PCR and overlap extension technique (SOE). In order to avoid the change of spatial structure and function after bimolecular attachment, polypeptide amino acid linker (Linker) was introduced between CD4 6 and CD5 5 and cloned to construct GPI anchored CD4 6 / CD5 5 Bluescript M13 cloning vector. Results The GPI anchored CD4 6 / CD5 5 chimeric cDNA with complete reading frame and correct junction was obtained. Conclusion This study laid the foundation for the development of a new generation of multifunctional and multi-target complement inhibitors