金黄色葡萄球菌表面蛋白的原核表达、纯化及免疫效应研究-运动人体科学专业论文.docxVIP

结论我们所获的金黄色葡萄球菌表面蛋白是可溶性的，较好地保持了其免疫 结论我们所获的金黄色葡萄球菌表面蛋白是可溶性的，较好地保持了其免疫 原性，实现了其在大肠杆菌中的高量表达：我们首次尝试将金葡菌的厂仃b和ona 基因连接起来，构建双价疫苗从而扩大了蛋白疫苗的保护范围。使之可以防治绝 大部分金葡菌株；动物实验表明，这些蛋白具有良好的体液免疫和细胞免疫应答 的效果；开发金葡菌ECMBPs重组疫苗具良好的应用前景。 关键词金黄色葡萄球菌：纤连蛋白结合蛋白；胶原蛋白结合蛋白；表达与 纯化；疫苗：免疫效应 Cloning，expressi9n，purification Cloning，expressi9n，purification and study on immunological effect ofStaph)7lococcus surface proteins Z／-／OUHong Abstract Staphylococcus number of infections of varying severity incomplicated wound infections which progress complicated septicaemia， osteomyelites，septic arthritis endocarditis．S is of the more bacterial species causing serious hospital—acquired infections．Metl,iicillin-resistant S oureus(MRSA)strains resist against most clinically useful antibiotics．As result of this，vancomycin is often the only effective antibiotic remaining．Moreover, mports from clinical studies have shown the existence of MRSA strains witll reduced sensitivity for vancomycin．The advent of Vancomysin-intermediate S ureus(VISAl strains have great panic which the infection of矗aureus couldn’t cure．This situation has also led to renewed interest in vaccination against Js_aureus infections complement antibiotic treatment and prophylactics． Recent research reveals that vaccines based recombinant staphylococcal extracellar-matrix-binding proteins(ECMBPs)are much more protective．Using PCR technology，we amplify thefnb and genes of S．aureus encode fibroneetin binding proteins fFnBP)and collagen binding proteins(Cna)，And cloning the expressioif pET-32a(+)fused with Trx·Tag coding sequence acquire the recombinant plasmid trx-fnb—pET一32a(+)，trx-cna-pET·32“+)，fnb—pET一32a(+)and cna-pET一32《+)．Induction by IPTG，we acquired the high expressed fusion protein Trx-Cna．Trx-FnBP(these proteins fur 58．8％、52．8％oftotal cellular proteins) and fusion protein Cna．In order get the bivalent protein vaccine，using overlapping primer extension PCR method，we linked the fnb and genes and cloning