青海血蜱幼蜱cDNA表达文库的构建及其保护性抗原基因的免疫学筛选  

Construction of cDNA expression library of Haemaphysalis qinghaiensis larval tick and immunoscreening of genes encoding protective antigens

作　　者：赵海平[1] 高金亮[1] 李有全[1] 樊瑞泉[1] 李金杰[1] 赵金国[1] 刘军龙[1] 殷宏[1] 罗建勋[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室

出　　处：《中国兽医科学》2007年第6期461-464,共4页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2006AA10A207);国家自然科学基金项目(30270992)

摘　　要：采用常规方法构建了青海血蜱幼蜱cDNA表达文库,初级文库的库容量为5.0×105PFU/mL,扩增后的库容量为8×109PFU/mL;用兔抗青海血蜱幼蜱阳性血清对该文库进行了免疫学筛选,对阳性噬菌体进行了亚克隆,提取质粒后转化宿主菌JM109,最终共得到幼蜱基因11个,分别命名为HqL1、HqL2、HqL3、HqL4、HqL5、HqL6、HqL7、HqL8、HqL9、HqL10、HqL11。测序分析表明,其中的6个为新基因。分析发现HqL7、HqL8、HqL9和HqL11具有较高的研究价值。A cDNA expression library of Haemaphysalis qinghaiensis larval tick was constructed. The capacity of the primary library was 5.0×10^5PFU/mL and the capacity of the amplified library was 8×10^9 PFU/mL. The cDNA library was immunoscreened by the rabbit serum against H. qinghaiensis larval tick, the positive pluques were subcloned, and their plasmids extracted and then transformed into Escherichia coli JM109. Eventually, 11 genes were obtained and designated HqL1, HqL2, HqL3, HqL4, HqL5, HqL6, HqL7,HqL8,HqLg, HqL10 and HqL11, respectively. Sequencing revealed that 6 of them represent new genes. Genes HqL7, HqL8, HqL9 and HqL11 are worth to be further studied.

关 键 词：青海血蜱 幼蜱 CDNA表达文库 免疫学筛选 

分 类 号：S852.746[农业科学—基础兽医学]