捻转血矛线虫ZJ株24000分泌排泄(ES)蛋白基因的克隆和序列分析

通过 RT- PCR方法 ,从捻转血矛线虫成虫总 RNA中扩增得到特异性片段 ,并把这一基因片段克隆到 p U C18克隆载体上 ,进行测序及同源性比较。序列分析可知 ,其核苷酸序列与国外已发表的 2 4 0 0 0分泌排泄抗原基因的同源性为 99.2 %。表明本试验成功提取了捻转血矛线虫成虫总 RNA,并用 RT- PCR方法克隆了 2 4 0 0 0分泌排泄抗原基因 ,为该基因的表达及其免疫学作用研究奠定了基础

Cloning and Sequence Analysis of 24 000 Excretory-secretory Antigen Gene of Haemonchus contortus ZJ Strain

A pair of primers were designed according to gene sequence of the published (24 000 ) excretory-secretory antigen in the database.Using RT-PCR the specific gene fragment was amplified from the total RNA extracted from the Haemonchus contortus adults,then the PCR product was linked to the pUC18 vector and the recombinant plasmid was verified by sequencing.The sequence result was analysized with BLAST software.The (24 000) excretory-secretory antigen gene has been successfully cloned using the RT-PCR.Sequence analysis showed that the gene is consistent to the (24 000) gene sequence in the database.After BLAST analysis the gene obtained is 99.2% identical to the (24 000) gene in the database.High quality total RNA was extracted from the Haemonchus contortus adults and cloned the (24 000) gene using PCR.The present work has laid a foundation for the future (24 000) gene expression and the related immunology research.