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Position: Home > Articles > Cloning and Prokaryotic Expression of Streptococcus Agalactiae Sip Gene Grass-Feeding Livestock 2016 (1) 36-41

无乳链球菌Sip基因的克隆与原核表达

作  者：
杨学云;王蒴;吴建勇;王登峰;李建军
单  位：
新疆畜牧科学院兽医研究所
关键词：
无乳链球菌;表面蛋白;Sip基因;克隆与表达
摘  要：
无乳链球菌表面免疫相关蛋白(Surface immunogenic proteins,Sip)具有高度保守性,是无乳链球菌疫苗研究的重要靶标。通过PCR技术扩增Sip基因,将其插入pET-22b载体构建重组质粒pET-22b-Sip,并对重组质粒进行双酶切、PCR鉴定及测序;转入BL21(DE3)中诱导表达,检测重组蛋白的大小及反应原性。结果表明,所扩增Sip基因大小为1 300bp,与GeneBank参考序列同源性为99.16%;蛋白分析表明,目的蛋白大小为50KDa,具有良好的反应原性,为无乳链球菌的免疫预防提供依据和基础。
译  名：
Cloning and Prokaryotic Expression of Streptococcus Agalactiae Sip Gene
作  者：
YANG Xue-yun;WANG Shuo;WU Jian-yong;WANG Deng-feng;LI Jian-jun;Institute of Veterinary Medicine, Xinjiang Academy of Animal Science;
关键词：
streptococcus agalactiae;;surface protein;;sip gene;;cloning and expression
摘  要：
Streptococcus Agalactiae surface immunogenic protein(Sip) is a highly conservative important target for the study of Streptococcus Agalactiae vaccine. The Sip gene was amplified by PCR and be inserted into vector p ET-22 b from a recombinant vector p ET-22b-Sip. After identified by PCR, restriction enzyme digestion and sequencing methods, the recombinant plasmid was transformed into BL21(DE3) and induced to express by IPTG. The size and reactionogenicity of recombinant protein was detected. The results showed that the sip gene was 1300 bp and was 99.16% in homology with reference sequence of Gene Bank. The recombinant protein was found to be about 50 KDa as expected and showed good reactionogenicity. This research provided the basis and foundation for the immunization against Streptococcus Agalactiae.

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