重组肿瘤血管生成抑制因子CTF的构建和特征分析  

Construction of CTF(Calreticulin Tumstatin Fusion) Gene and Its Bioinformatics Analysis

作　　者：顾取良[1] 张添元[2] 

机构地区：[1]广东药学院基础学院,广州510006 [2]中山大学生命科学学院,广州510275

出　　处：《基因组学与应用生物学》2015年第3期476-481,共6页Genomics and Applied Biology

基　　金：广东药学院科研启动基金项目(2005JCB16);国家自然科学基金(31100852)共同资助

摘　　要：为获得一种更高效的重组肿瘤血管生成抑制因子,利用重叠延伸PCR技术将来自钙网蛋白(calreticulin)和肿瘤抑素(tumstatin)的抗血管生成活性片段连接,构建融合基因CTF(calreticulin tumstatin fusion)并克隆至表达载体p ET-15b。运用生物信息学软件对其分子结构的生物学特征进行分析发现,CTF融合基因编码94个氨基酸,分子量为10.72 k D,理论等电点为7.68,利于基因工程表达;连接肽段部位呈中性且柔性良好,不影响两端活性区域原来的二级结构和融合蛋白保守结构域。研究结果表明,本研究构建的CTF能够保留钙网蛋白活性片段和肿瘤抑素活性片段各自空间结构,适合融合蛋白生物活性的发挥。To obtain a more potent recombinant angiogenesis inhibitor, bioactive anti-angiogenic fragments from calreticulin and tumstatin were generated by polymerase chain reaction(PCR) respectively and then were linked with GSGGSG bridge by splicing overlapping extension(SOE), resulting in a synthetic fusion gene CTF, which was further cloned into p ET-15 b vector. Bioinformatics approaches were used to predict the properties including the flexibility and hydrophobicity, secondary structural features and conserved domains for recombinant CTF fusion protein. Recombinant CTF contains 94 amino acids, with molecular weight of 10.72 k D and theoretical p I at7.68. Structure analysis showed the fusion protein CTF maintained original domains of bioactive fragments from calrecticulin and tumstatin, with a linker of high flexibility. We would have a conclusion that the construction of CTF gene might provide a rational approach for designing novel recombinant angiogenesis inhibitors with potent biological activity.

关 键 词：血管生成抑制因子 钙网蛋白 肿瘤抑素 生物信息学 

分 类 号：R730.2[医药卫生—肿瘤]