摘要
根据带科绦虫8ku蛋白基因序列设计引物,采用RT-PCR方法对牛带绦虫六钩蚴总RNA进行扩增,扩增产物经胶回收试剂盒纯化,与pGEM-T Easy载体连接,转化大肠杆菌JM109感受态细胞,筛选含有阳性重组DNA的克隆,对阳性克隆进行测序。使用DNAStar和MAGE 4生物学软件对基因克隆进行序列分析和进化树分析。结果显示,成功克隆了5种牛带绦虫8ku蛋白基因家族新成员,cDNA序列完整开放阅读框的大小为258bp,TSA8ku-1cDNA为优势克隆。序列同源性分析表明,各cDNA克隆氨基酸序列之间的差异性为1.2%～14.2%,与猪带绦虫诊断抗原TsRS1群(组)成员同源,相似性为85.9%～88.9%。由此可以推测,克隆的5种8ku蛋白基因家族成员可能是牛带绦虫病的重要诊断抗原候选基因。
Complementary DNAs（cDNAs）of taeniid cestode 8 ku protein gene family were ampl ified from total RNA of hatched and activated Taenia saginata on cospheres using RT-PCR.The purified PCR products were inserted into pGEM-T Easy vector,and then transf ormed into competent Escherichia coli JM109.Positive clones containing the recombinant plasmids were identified by PCR and e nzyme restriction analysis and then sequenced.Sequences were analyzed using DNAS tar and MAGE 4 softwares.In results,5 cDNA sequences were obtained with a 258 bp of open reading frame.T hese sequences belonged to the T.saginata 8 ku gene family with 1.2% to 14.2% amino acid residue divergence,and the TSA8ku-1 was a dominant cl one.Comparison with relative taeniid cestode 8 ku protein sequences indicated 8 5.9% to 88.9% similarity of amino acids between the TSA8ku proteins and diagnos tic antigen TsRS1 group from Taenia solium,suggesting that the T SA8ku proteins should be candidate and important immunodiagnostic antigens for b ovine cysticercosis.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2010年第9期881-885,共5页
Chinese Veterinary Science
基金
国家科技支撑计划项目(2007BAD40B00)
公益性行业(农业)科研专项(200903036-07)
家畜疫病病原生物学国家重点实验室自主课题(SKLVEB2008ZZKT014)
