长角血蜱甘肃株肌钙蛋白基因P27/30的克隆和序列分析  

Cloning and sequencing of the P27/30 gene of Gansu strain of haemaphysalis longicomis

作　　者：田占成[1] 刘光远[1] 李知新[1] 谢俊仁[1] 王路[1] 张林[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/甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国预防兽医学报》2007年第6期413-416,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技基础条件平台项目(2005DKA21205-3);甘肃省自然科学基金项目(32S041-A25-035)

摘　　要：参照GenBank中长角血蜱致病性Okayama株肌钙蛋白P27/30基因的核苷酸序列,设计合成一对引物,从本实验室保藏的干净长角血蜱卵中快速提取总RNA,通过RT-PCR扩增出607 bp的肌钙蛋白基因,将其克隆于PGEM-T-Easy载体,对其进行序列分析,并推导出氨基酸序列,将这一序列与国内外已发表的长角血蜱肌钙蛋白基因进行比较分析。结果表明本实验室保藏的长角血蜱甘肃株与上述已发表的Okayama株核苷酸序列同源性为92.1%,氨基酸同源性为86%。长角血蜱甘肃株与Okayama株、四川孤雌生殖株的发育有一定差异。经RT-PCR分析表明,该基因在长角血蜱的卵、幼蜱、若蜱和饥饿成蜱这几个阶段均有表达。The P27/30 gene of haemaphysalis longicomis Gansu strain was amplified by RT-PCR from genomic RNA and cloned into pGEM-T plasmid vector and sequenced. Comparison and analysis of the nucleotide sequence and amino acid sequence with those of other 6 strains previously published were performed using DNAStar software. The results showed that the deduced amino acid sequences of these strains shared 46 %-86 % homology. The Gansu strain and Okayama strain shared homologies of 92.1% at the nucleotide level and 86 % at amino acid level. The results indicated that the Gansu strain may differ from other sWains in their developmental stages. The P27/30 gene could be detected in each developmental stage of haemaphysalis longicomis.

关 键 词：长角血蜱 肌钙蛋白基因P27/30 克隆 序列分析 

分 类 号：Q781[生物学—分子生物学] S852.746[农业科学—基础兽医学]