摘要
为了研究家蝇几丁质结合蛋白(Md-CBPⅠ)在家蝇防御体系中的相关活性,采用PCR技术,对从鸡源沙门氏菌诱导家蝇幼虫构建的抑制性消减文库(SSH)中筛选到的家蝇幼虫几丁质结合蛋白Ⅰ基因(Musca domestica chitin binding proteinⅠ,Md-CBPⅠ)进行扩增,并成功构建了重组表达质粒p ET-32a-Md-CBPⅠ,于大肠杆菌BL21(DE3)中得以高效表达,纯化并获得了MdCBPⅠ融合蛋白。进一步对该蛋白的亲和活性进行了研究,发现Md-CBPⅠ融合蛋白对几丁质以及纤维素均有一定的结合作用,且其对几丁质的结合作用相对较好。试验为家蝇几丁质结合蛋白生物学活性和免疫学活性的研究奠定了基础。
In order to research the activities of Md - CBP I in musca domestica defence system, Md - CBP I was screened from suppression subtractive library （SSH） in Musca domestica larvae induced by Salmonella pullorum of chickens and also amplified by PCR techniques. The recombinant expression plasmid of pET - 32a - Md - CBP I was constructed successfully and expressed in E. Coli BL21 （DE3）. fusion further more, the preliminary study was made on the affinity of which reveal protein was obtained by purification, ed that the fusion protein has binding affinity of both chitin and cellulose, and the affinity of chitin was stronger than cellulose. This research laid a foundation for further research on biological activity and immunological activity of Md - CBP I
出处
《中国兽药杂志》
北大核心
2016年第1期8-13,共6页
Chinese Journal of Veterinary Drug
基金
国家自然科学基金资助项目(31572574,31502121)
吉林省世行贷款农产品质量安全项目(2011-Y05)
关键词
家蝇
几丁质结合蛋白
克隆
原核表达
Musca domestica
chitin binding protein
clone
prokaryotic expression
