多房棘球绦虫重组BCG-EmⅡ／3-10疫苗的构建  

CONSTRUCTION OF RECOMBINANT BCG-EM Ⅱ/3-10 VACCINE OF ECHINOCOCCUS MULTILOCULARIS

作　　者：郭鄂平[1] 陈林[2] 张光玉[1] 王绍基[1] 宋明华[1] 

机构地区：[1]郧阳医学院寄生虫学教研室,十堰442000 [2]郧阳医学院附属太和医院药学部

出　　处：《现代预防医学》2009年第4期714-716,共3页Modern Preventive Medicine

基　　金：湖北省教育厅科研基金资助项目(2004D011)

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。[Objective] To construct recombinant BCG-EmⅡ/3-10 vaccine of Echinococcus muhilocularis. [_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-Myeobacterium shuttle plasmid pBCG to construct recombinant pBCG-Em Ⅱ/3-10. The plasmid was introduced into BCG by electroporation to construct rBCG-Em Ⅱ/3-10 vaccine. The vaccine was cultured for 3 weeks in Sauton medium with 12mg/L HgCl2. [ Results] It was successful that the construction of pBCG-Em Ⅱ/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-Em Ⅱ/3-10 vaccine of E. muhilocularis is successfully constructed and lays foundation for exploitation and utilization of this vaccine.

关 键 词：多房棘球绦虫 重组BCG-EmⅡ/3-10疫苗 构建 鉴定 

分 类 号：R383.3[医药卫生—医学寄生虫学]