中文摘要：

目的构建多房棘球绦虫重组质粒pBCG-Em14-3-3．并转化BCG进行表达。方法超声粉碎泡球蚴组织提取总RNA，通过RT-PCR扩增Em14-3-3抗原编码基因；将该扩增产物定向克隆到大肠杆菌-分枝杆菌穿梭表达载体pBCG，构建重组质粒pBCG-Em14-3-3；电穿孔法转化BCG，构建多房棘球绦虫重组BCG-Em14-3-3。免疫印迹分析pBCG-Em14-3-3的表达产物。结果RT-PCR成功扩增出779bp的Em14-3-3抗原编码基因；双酶切证实Em14-3-3抗原编码基因成功插入pBCG中；PCR证实rBCG-Em14-3-3构建成功；免疫印迹分析发现pBCG-Em14-3-3的表达产物在相对分子质量（Mr）约为27×10^3处有明显的目的蛋白表达条带，且能被活动性泡球蚴病鼠血清特异识别。结论成功构建了多房棘球绦虫重组BCG-Em14-3-3。

英文摘要：

To construct the recombinent plasmid pBCG-Em 14-3-3 of E. multilocularis and to explore its expression in BCG. the total RNA was extracted from alveolar hydatid cyst by ultrasound-breaking, and Em 14-3-3 antigen gene was amplified by RT-PCR from the total RNA. This PCR product was cloned into Escherichia coli-Mycobacterium shuttle plasmid pBCG to construct the recombinant plasmid pBCG- Em 14-3-3. The recombinant plasmid was electroporated into BCG to construct rBCG- Ern 14-3-3,and the expression product of rBCG- Em 14-3-3 was identified by western blotting. It is dEmonstrated that the 779bp Ern 14-3-3 gene was successfully amplified by RT-PCR and cloned into pBCG by restriction analysis. In addition rBCG- Ern 14-3-3 was successfully constructed by PCR, the expression product had obviously target protein in Mr 27 × 10^3 ,which could be recognized by sera from mice infected with AE. Thus rBCG- Em 14-3-3 of E. multilocularis is successfully constructed.