厚壳贻贝粘附蛋白十肽重复序列的表达及应用

 利用PCR扩增 Mcfp-1（M. coruscus foot protein-1）基因的12个十肽重复序列粘附功能片段（Mcfp~1 1~12）并连接到pGEX-4T1表达载体中，在大肠杆菌BL21(DE3)中诱导表达.纯化的多肽产物经凝血酶处理切除GST标签，获得Mcfp-1 1~12功能肽段，最后用酪氨酸酶对该产物进行修饰.通过材料表面包被、石英晶体微天平（QCM, quartz crystal microbalance）分析、细胞粘附和细胞毒性等实验,研究了该表达产物作为生物粘合剂的粘附特性.结果显示，重组表达产物Mcfp-1 1~12在多种材料表面的包被能力与Cell-TakTM（天然提取的贻贝粘附蛋白混合物）相当，甚至更佳；对细胞的粘附能力与Cell-TakTM相当；用HeLa细胞和293T细胞进行的MTT实验未发现细胞毒性.上述结果表明,Mcfp-1 1~12作为生物医用粘合剂具有潜在应用价值；同时，基因重组技术可以为制备新型海洋贻贝粘附蛋白防水生物粘合剂提供有效的手段.本研究为临床使用更优质的生物医用粘合剂提供了理论依据.

Expression and Application of Recombinant Decapeptide Repeats of Mytilus coruscus Foot Protein as bioadhesive

The decapeptidyl repeats from M.coruscus foot protein-1(Mcfp-1) was amplified by PCR and cloned into pGEX-4T1 expression vector.The recombinant Mcfp-1 fragment 1-12 GST fusion was expressed in E.coli BL21(DE3) and purified with glutathione-Sepharose 4B.Thrombin was used to remove GST to obtain the recombinant the Mcfp-1 1-12 peptide,and the adhesive bioactivity was characterized by tyrosinase modification,surface coating analysis,QCM(quartz crystal microbalance),and cell adhesion and cytotoxicity analysis.T...