【摘要】 [目的]构建多房棘球绦虫重组BCG-EmⅡ/3-10疫苗。[方法]超声粉碎泡球蚴组织提取总RNA,通过RT-PCR扩增EmⅡ/3-10抗原编码基因,将该基因定向克隆到大肠埃希菌-分枝杆菌穿梭表达载体pBCG,构建重组质粒pBCG-EmⅡ/3-10,电穿孔法将该质粒导入BCG构建多房棘球绦虫重组BCG-EmⅡ/3-10疫苗。将该疫苗接种到含12mg/LHgCl2的罗氏培养基筛选培养3周。[结果]重组pBCG-EmⅡ/3-10质粒经酶切证实构建成功,重组BCG-EmⅡ/3-10疫苗能在含12mg/LHgCl2的罗氏培养基上生长繁殖。[结论]成功构建了多房棘球绦虫重组BCG-EmⅡ/3-10疫苗,为疫苗的开发和利用打下了基础。更多还原

【Abstract】 [Objective] To construct recombinant BCG-EmⅡ/3-10 vaccine of Echinococcus multilocularis.[Methods] The total RNA was extracted by ultrasound-breaking from alveolar hydatid cyst.The EmⅡ /3-10 antigen gene was amplified by RT-PCR from the total RNA and then was cloned into E.coli-Mycobacterium shuttle plasmid pBCG to construct recombinant pBCG-EmⅡ /3-10.The plasmid was introduced into BCG by electroporation to construct rBCG-EmⅡ /3-10 vaccine.The vac-cine was cultured for 3 weeks in Sauton medium with 12mg /L HgCl2.[Results] It was successful that the construction of pBCG-EmⅡ /3-10 was confirmed by restrictive endonuclease digestion.The rBCG-Em Ⅱ /3-10 vaccine could reproduce in Sauton medium with 12mg /L HgCl2.[Conclusion] The rBCG-EmⅡ /3-10 vaccine of E.multilocularis is successfully con-structed and lays foundation for exploitation and utilization of this vaccine.更多还原