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微小隐孢子虫子孢子表面抗原CP23融合蛋白在大肠杆菌的表达
引用本文:何宏轩,张西臣,欧阳红生,尹继刚,李吉平,李建华,杨举. 微小隐孢子虫子孢子表面抗原CP23融合蛋白在大肠杆菌的表达[J]. 中国人兽共患病杂志, 2003, 19(3): 52-54
作者姓名:何宏轩  张西臣  欧阳红生  尹继刚  李吉平  李建华  杨举
作者单位:解放军军需大学军事兽医系,解放军军需大学军事兽医系,解放军军需大学军事兽医系,解放军军需大学军事兽医系,解放军军需大学军事兽医系,解放军军需大学军事兽医系,解放军军需大学军事兽医系 长春,130062;河南职业技术师范学院动物科学系,长春,130062,长春,130062,长春,130062,长春,130062,长春,130062,长春,130062
基金项目:吉林省杰出青年基金 (2 0 0 0年度 )
摘    要:目的 构建微小隐孢子虫子孢子表面抗原CP2 3的原核表达载体 ,并且在大肠杆菌中表达。方法 用HindⅢ和EcoRⅠ酶分别从 pET - 2 8a(+)和 pMD18-T - 2 3质粒中酶切得到线性片段和CP2 3基因片段 ,然后用T4酶连接 ,构建重组的CP2 3的原核表达载体 ,在大肠杆菌BL2 1(DE3)中表达 ,并用SDS -PAGE、ELISA和Westernblotting进行鉴定。结果 构建了CP2 3的原核表达载体 ,得到了一分子量约为 14kDa的融合蛋白 ,占大肠杆菌总蛋白的 36 %。结论 CP2 3融合蛋白在大肠杆菌中得到了高效表达

关 键 词:隐孢子虫  CP23  大肠杆菌  表达  
文章编号:1002-2694(2003)03-0052-03
收稿时间:2003-03-20
修稿时间:2001-11-04

Expression of CP23 fusion protein on sporozoites of Cryptosporidium parvum in E. coli
HE Hongxuan,ZHANG Xichen,OUYANG Hongsheng,et al. Expression of CP23 fusion protein on sporozoites of Cryptosporidium parvum in E. coli[J]. Chinese Journal of Zoonoses, 2003, 19(3): 52-54
Authors:HE Hongxuan  ZHANG Xichen  OUYANG Hongsheng  et al
Abstract:Aims To construct expression vector of CP23 on sporozoites of Cryptosporidium parvum and express it in E coli Methods A linear fragment of pET-28a(+) and CP23 gene of C parvum were obtained from the plasmids of pET-28a(+) and pMD18-T-23 with Hind Ⅲ and EcoRⅠdisgested The expression vector of CP23 was constructed by T 4 ligase and transformed into BL21(DE3) strain cells of E coli for expression The expression products were analyzed by SDS-PAGE,ELISA and Western blotting Results The expression vector of CP23 was constructed Distinct protein band with a molecular weight of 14 kDa was detected on SDS-PAGE and its antigenicity was confirmed by ELISA and Western blotting Conclusion Efficient expression of CP23 fusion protein has been achieved in E coli
Keywords:Cryptosporidium parvum  CP23  E coli  gene expression
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