空肠弯曲菌penner o:19 cjaA基因的克隆表达

目的克隆空肠弯曲菌penner o:19 cjaA基因,构建其原核表达载体,诱导表达及纯化蛋白,为基因工程疫苗的研制打下基础。方法扩增空肠弯曲菌penner o:19 cjaA基因,构建pGEM-T-cjaA克隆载体,以EcoRI和XhoI为酶切位点构建pGEX-4T-1-cjaA原核表达载体。表达载体转化E.coli BL21后诱导表达,SDS-PAGE鉴定表达产物,用Glutathione Sepharose4B纯化重组蛋白。结果成功构建pGEX-4T-1-cjaA表达载体,并获得浓度为10μg/μl的rGST-CjaA重组蛋白。结论成功构建pGEM-T-cjaA克隆载体及pGEX-4T-1-cjaA表达载体,获得重组蛋白rGST-CjaA,为研究空肠弯曲菌疫苗打下基础

Construction of the prokaryotic express vector of cjaA gene of campylobacter jejuni

目的克隆空肠弯曲菌PEnnEr O:19 CJAA基因,构建其原核表达载体,诱导表达及纯化蛋白,为基因工程疫苗的研制打下基础。方法扩增空肠弯曲菌PEnnEr O:19 CJAA基因,构建PgEM-T-CJAA克隆载体,以ECOrI和XHOI为酶切位点构建PgEX-4T-1-CJAA原核表达载体。表达载体转化E.COlI bl21后诱导表达,SdS-PAgE鉴定表达产物,用gluTATHIOnE SEPHArOSE4b纯化重组蛋白。结果成功构建PgEX-4T-1-CJAA表达载体,并获得浓度为10μg/μl的rgST-CJAA重组蛋白。结论成功构建PgEM-T-CJAA克隆载体及PgEX-4T-1-CJAA表达载体,获得重组蛋白rgST-CJAA,为研究空肠弯曲菌疫苗打下基础。Objective To construct prokaryotic expression vector carrying campylobacter.jejuni penner o:19 cjaA gene,express it in E.coli and purify protein.Methods The cjaA gene were amplified by PCR.The clone vector pGEM-T and expression vector pGEX-4T-1 with inserted cjaA gene were constructed.The target protein was expressed in E.coli BL21.The protein was identified and analyzed by SDS-PAGE.The target protein was purified by Glutathione Sepharose4B.Results Constructed the expression vector pGEX-4T-1-cjaA,purified the recombinant protein.Conclusion The recombinant plamid pGEM-T-cjaA and pGEX-4T-1-cjaA have been successfully constructed and the GST-CjaA protein is purified,which lays the foundation for the further development of C.jejuni genetic engineering vaccine.国家自然科学基金资助项目(30471919);国家高技术研究发展计划(2006AA02A208);(863计划2006AA02A2080

基于pH的反馈补料方法在谷氨酸发酵中的应用

为简化谷氨酸发酵补料工艺,提出了一种新型的基于pH的补料方式。考察谷氨酸发酵过程中氨消耗量(x)和糖消耗量(y)发现,两者之间存在较好的线性关系(y=7.4744x,R2=0.9989),以此为pH反馈补料工艺中补料液中葡萄糖与氨的混合比例,能较好地将谷氨酸发酵过程中葡萄糖浓度稳定在12~21 g/L。比较恒定葡萄糖浓度补料工艺与pH反馈补料工艺发现,采用pH反馈补料工艺进行发酵,葡萄糖转化率、谷氨酸产酸速率分别提高了9.06%和17.5%左右,同时发酵周期缩短2 h以上