摘要
目的在人大肠杆菌中表达人ＣＤ８０胞外区蛋白质。方法采用ＰＣＲ扩增人ＣＤ８０胞外区基因ｃＤＮＡ，先后经Ｋｌｅｎｏｗ和ＨｉｎｄⅢ酶切并纯化，再定向克隆至高效表达载体ｐＫｐＬ－３ａ中，构建成ｐＫｐＬ－ＣＤ８０。转化大肠杆菌ｐｏｐ２１３６后，温度诱导表达，再用Ｗｅｓｔｅｒｎｂｌｏｔ鉴定之并检测其活性。结果转化菌表达出分子量为２４０００的蛋白质，免疫学鉴定为ＣＤ８０，并证实有一定生物学活性。结论大肠杆菌中可以成功表达出具有生物学活性的ＣＤ８０，为进一步研究和开发ＣＤ８０打下了基础。
Objective To express and identify the extracellular region of human CD80 protein in E.coli . Methods The extracellular region cDNA of human CD80 was amplified with PCR followed by digestion with Klenow fragment and Hind Ⅲ,and then subjected to unidirectionally cloning into pKpL 3a.resulted pKpL CD80 was transformed into E.coli . pop2136 and expressed by induction at 42℃ for 4 hours.The expressed protein was identified by Western blor with McAb anti human CD80 derived from mice,and its biological activity was tested.Results The extracellular region of human CD80 protein was successfuly expressed in E.coli . pop2136,and was identified by Western blot.Its biological activity was also verified.Conclusions Expression of this recombinant protein of the extracellular region of human CD80 will provide the possibility for preparation of CD80 on large scale,then make it possible to further study and exploit the function of CD80.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1998年第3期246-249,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金
霍英东青年教师基金
关键词
人CD80
基因
表达
大肠杆菌
Human CD80 Gene Expression\ \ Escherichia coli
