猪旋毛虫不同发育时期虫体49ku ES抗原基因的克隆与序列分析  

Cloning and sequence analysis of 49 ku ES antigen gene of Trichinella spiralis at different developmental stages

作　　者：路义鑫[1] 宋铭忻[1] 王洪斌[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2006年第11期894-897,共4页Chinese Veterinary Science

基　　金：霍英东青年教师基金项目(91034);中国博士后科学基金项目(2004035160);哈尔滨市学科后备带头人基金项目(2004AFXXJ051)

摘　　要：根据GenBank中已登录的猪旋毛虫49 ku ES抗原基因序列设计了1对引物,用SDS裂解液配合蛋白酶K方法从猪旋毛虫不同发育时期(成囊前期幼虫、肌幼虫、成虫)虫体中提取总RNA,用RT-PCR方法扩增49 ku ES抗原基因,将目的基因定向克隆入pMD18-T载体,转化大肠埃希氏菌TG1。用PCR、限制性内切酶EcoRⅠ和BamHⅠ进行单、双酶切鉴定,阳性质粒的测序结果表明,成功克隆了猪旋毛虫不同发育时期虫体49 ku ES抗原基因。序列分析结果显示:49 kuES抗原基因大小为948 bp,基因的保守性很强,序列同源性比较高,不同发育时期之间的差异很小。A pair of primers for the 49 ku ES antigen gene of Trichinella spiralis was designed according to the published sequence in GenBank （accession number M64242）. The total RNAs were extracted from different developmental stages of T. spiralis（including pre-encysted larvae, muscle larvae and adult worms） by the SDS spallation liquid, together with Protease K, respectively. The genes encoding 49 ku ES antigen were amplified by RT-PCR. The target genes were directly cloned into the pMD18-T Vector, and then transferred into Escherichia coli strain TG1. The recombinant plasmids were identified by enzymatic digestion with EcoR I and BamH I and PCR amplification. The positive plasmids were subjected to sequencing. The 49 ku ES antigen genes at different developmental stages were cloned successfully. Sequence analysis indicated that the 49 ku ES gene was 948 bp in length and was conserved, and the homology rates among the sequences was quite high at different developmental stages.

关 键 词：旋毛虫 49 KU ES抗原基因 克隆 序列分析 

分 类 号：S852.731[农业科学—基础兽医学] Q785[农业科学—兽医学]