捻转血矛线虫Hc38保守结构域所在基因片段的真核表达载体构建  

Construction of an Eukaryotic Expression Vector of Hc38 Gene of Conserved Domain of Haemonchus Contortus

作　　者：郑金海[1] 段正秀[2] 荣光[3] 薄新文[1] 李娜[1] 何立雄[1] 

机构地区：[1]新疆兵团绵羊繁育生物技术重点实验室,新疆石河子832000 [2]石河子大学生命科技学院,新疆石河子832000 [3]中国热带农业科学院,海南儋洲571737

出　　处：《动物医学进展》2009年第2期22-25,共4页Progress In Veterinary Medicine

基　　金：新疆兵团科技局社会发展项目(2006GG31)

摘　　要：根据捻转血矛线虫Hc38基因产物(登录号AY749124)保守结构域所在核酸序列设计1对引物,该引物包含Kozak序列、HindⅢ酶切位点、XbaⅠ酶切位点。以含有捻转血矛线虫Hc38基因保守结构域片段的pMD19-T为模板,经PCR扩增获得Hc38基因保守结构域片段,用HindⅢ、XbaⅠ双酶切该片段,回收后将此基因片段克隆至相同酶切回收后的真核表达载体pcDNA3.1(+)中,获得重组质粒pcD-NA3.1-Hc38。经酶切分析、序列测定和PCR鉴定,证实了重组质粒的正确性。According to Hc38 gene（GenBank number： AY749124）of Haemonchus contortus, a pair of primers with Kozak sequence, HindⅢ and Xba Ⅰ sits were densigned. The fragment in conserved domain was amplified by PCR from pMD19T-Hc38. The PCR product was digested with HindⅢ and Xba Ⅰ ,then inserted into pcDNA3.1 （＋） between Hind Ⅲ and Xba Ⅰ sites to generate an expression plasmid pcDNA3.1 Hc38. The recombinant plasmid was identified by enzyme digestion,PCR and sequence. The eukaryotic ex pression plasmid pcDNA3.1-Hc38 was successfully constructed.

关 键 词：捻转血矛线虫 Hc38基因 真核表达载体 KOZAK序列 

分 类 号：S855.9[农业科学—临床兽医学] S852.731[农业科学—兽医学]