中文摘要：

贻贝（Mytilus galloprovincialis）足丝中黏附蛋白,是一种黏合强度高、生物相容性好、优质的生物黏合剂,可以应用于医学、表面化学、海洋工程等领域,其中富含DOPA（多巴,3,4-二羟基苯丙氨酸）的贻贝足丝蛋白5（Mytilus galloprovincialis foot protein type 5,Mgfp-5）显著表现出此特性。天然获取Mgfp-5含量低,易固化,纯化困难,越来越多学者尝试基因工程获得黏蛋白。文中构建重组质粒p ET28a-mgfp,在大肠杆菌（Escherichia coli）BL21（DE3）中诱导表达出重组蛋白Mgfp-5。为得到高质量的Mgfp-5蛋白,优化了Mgfp-5蛋白诱导表达参数：菌液OD_（600）=0.8,诱导剂异丙基-β-D巯基半乳糖苷（IPTG）0.8 mmol/L,37℃,诱导8h;优化镍离子亲和层析纯化蛋白Mgfp-5最佳条件为：Elution Buffer的p H为7.0,500 mmol/L咪唑。Western Blot鉴定到重组Mgfp-5可以特异性表达。该研究为得到大量Mgfp-5蛋白奠定了基础,为加速黏蛋白的黏附机理及生物医学黏合剂的开发提供参考。

英文摘要：

The byssus of Mytilus galloprovincialis produce and secrete specialized adhesives Mytilus galloprovincialis foot protein type 5（ Mgfp-5） which has significant adhesive ability and biocompatibility and which is rich in high DOPA. It can be widely used in medical,chemical surface and ocean engineering or other fields.But the natural extraction of Mgfp-5 resulted in very little purified protein since this process is labor-intensive and inefficient,and these issues restrict its application. To obtain enough Mgfp-5 for the further study,gene fragments mgfp-5 were amplified and cloned it into the prokaryotic expression vector p ET28 a,then transformed it into Escherichia coli（ E. coli） BL21（ DE3） expression system. The expression of Mgfp-5 protein was induced by isopropyl-β-d-thiogalactoside（ IPTG） and SDS-PAGE was used to identify Mgfp-5. The recombinant plasmid p ET28a-mgfp was constructed correctly and the recombinant protein was expressed successfully. Thefactors including bacteria biomass,temperature,time and concentration of IPTG were optimized before induction. Recombinant protein was purified using Ni-NTA Purification System under native conditions. The optimum conditions for the induced expression of recombinant protein were determined as follows： the OD_（600） of bacterial before induction with 0. 8,the IPTG concentration of 0. 8 mmol / L,the induction temperature of 37℃and the induction time of 8 h. Recombinant protein was eluted with Native Elution Buffer（ p H 7. 0） containing 500 mmol / L imidazole. The specificity of the recombinant protein was confirmed using western blots. It can provide the basis for the Mgfp-5 manufacture,in-depth theoretical research and practical application. It can also provide a new way to develop a bioadhesive in medical or underwater environments.