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

Expression of CP23 fusion protein on sporozoites of Cryptosporidium parvum in E.coli

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

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