摘要
根据Gen Bank中已发表的序列(M64242)为参考设计引物,用RT-PCR方法从6个旋毛虫隔离种的成囊前期幼虫中扩增出了49 ku ES抗原基因,将目的基因定向克隆入pMD18-T载体,转化大肠杆菌TG1。用PCR、限制性内切酶EcoR I和BamHI进行单、双酶切鉴定,阳性质粒测序。成功克隆了6个旋毛虫隔离种成囊前期幼虫49 ku ES抗原基因,序列分析结果显示:49 kuES抗原基因的大小为948 bp,基因的保守性很强,序列同源性比较高,不同隔离种之间的差异很小。
A pair of primers for the 49 ku es gene of Trichinella spiralis were designed according to the published sequence in GenBank （accession number M64242）. The gene encoding 49 ku ES antigen were specifically amplified by RT-PCR. The target gene was directly cloned into the pMD18-T Vector, and then transformed 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 in six Trichinella isolates of pre-encysted larvae were cloned successfully. Sequence analysis indicated that the 49ku es gene was 948bp in length and was conserved, and the homology rates of the sequences were quite high among different Trichinella isolates.
出处
《中国兽医杂志》
CAS
北大核心
2007年第7期7-10,共4页
Chinese Journal of Veterinary Medicine
基金
哈尔滨市学科后备带头人基金(2004AFXXJ051)
霍英东青年教师基金(91034)
中国博士后科学基金(2004035160)
关键词
旋毛虫
成囊前期幼虫
克隆
序列分析
Trichinella spiralis
Pre-encysted larvae
Cloningl Sequence analysis
作者简介
路义鑫（1968-），男，副教授，博士，主要从事寄生虫病及寄生虫分子生物学研究。 电话：0451-55190729
通讯作者：王洪斌，宋铭忻.E-mail：songmx@neau．edu．cn
