肝片吸虫保护性抗原基因在卡介苗中的表达

Expressing F.hepatica Protective antigenic gene in BCG

用EcoRⅠ酶切含肝片吸虫保护性抗原基因FH3的重组质粒pUC18/FH3,回收FH3(约1.0kb)片段并测定其碱基序列 .FH3片段以正确相位插入到E .coli 分枝杆菌穿梭表达质粒载体pMV2 61中 ,构建含有肝片吸虫保护性抗原基因的重组穿梭质粒 .通过电转移法转化卡介苗 ,斑点杂交实验证明 ,重组卡介苗中含有此抗原基因 .热诱导FH3基因在卡介苗中的表达 ,并对基因表达产物进行SDS PAGE分析 结果显示 ,表达产物相对分子量为 2 2kD .

The protective antigenic gene FH3 of F.hepatica was cut out from recombinant plasmid pUC18/FH3 using EcoR Ⅰ ,sequenced and then inserted in the E.coli mycobacteria shuttle expression plasmid vector pMV261 in correct reading frame to construct a recombinant shuttle plasmid including the protective antigenic gene of F.hepatica named pMV261 FH3. Using electroporation, the recombinant plasmid was introduced into BCG. By dot blotting it was confirmed that the pMV261 FH3 had been inserted in recombinant BCG chromosome. Under heat induced(45℃), the expression recombinant protein was analyzed by SDS PAGE and its relative molecular mass is 22kD. The results may lay a foundation for constructing recombinant BCG vaccine against F.hepatica.