一种新型融合蛋白(RGD)3/tTF的基因表达与活性分析  

Gene Expression and Activities Analysis of a New Fusion Protein (RGD)3/tTF

作　　者：颜江华[1] 杨桂旺[1] 王阶平[1] 吴娜[1] 庄国洪[1] 

机构地区：[1]厦门大学医学院抗癌研究中心,厦门361005

出　　处：《生物工程学报》2007年第3期409-412,共4页Chinese Journal of Biotechnology

基　　金：福建省自然科学基金(No.C0410004);厦门大学科技创新基金资助(No.XDKJCX20053026)。~~

摘　　要：为了发展一种新型的融合蛋白(RGD)3/tTF用于肿瘤血管的选择性栓塞治疗,利用PCR技术重组(RGD)3/tTF融合基因,克隆于pET22b(+)载体,表达于E.coliBL21(DE3)。用镍柱纯化融合蛋白。凝血实验与FⅩ活化实验检测融合蛋白tTF组分的活性。间接ELISA分析(RGD)3/tTF与αvβ3的特异结合能力。pET22b(+)/(RGD)3/tTF重组质粒成功获得并表达于E.coliBL21(DE3)。纯化蛋白(RGD)3/tTF能有效诱发血液凝固,活化FⅩ。(RGD)3/tTF与αvβ3的特异结合能力比RGD/tTF提高了32%。新型融合蛋白(RGD)3/tTF已在E.coli系统成功表达,表达蛋白保持tTF的活性并显示比RGD/tTF更高的与αvβ3的结合能力。To develop a new fusion protein （RGD）3/tTF for the therapy of the selective thrombosis of tumor blood vessels. The fused gene （RGD）3/tTF was reconstructed by PCR, was cloned into vector pET22 b（ ＋ ）, and expressed in E. coli BL21 （ DEs ）. The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was detected by clotting assay and F X activation assay. The specific binding of （RGD）3/tTF to avβ3 was analyzed by indirect ELISA. The recombinant plasmid pET22 b（ ＋ ）/（RGD） 3/tTF was obtained and expressed in E. coil BI21 （DEs）. The purified fusion protein could induce blood coagulation, activiate F X. The ability of （RGD）3/tTF binding specifically to avβ3 was increased by 32%, compared with RGD/tTF. A new fusion protein （RGD）3/tTF was successfully expressed in E. coil B[21 （DEs）. The expressed proteins retained tTF activity and showed a higher binding to avβ3 than that of RGD/tTF.

关 键 词：TTF RGD 融合蛋白 凝血 

分 类 号：R346[医药卫生—基础医学]