细粒棘球蚴线粒体ND1基因的克隆与序列研究

Cloning and Sequence Analysis of the Partial ND1 Gene within Mitochondrial DNA of Echinococcus granulosus

为了研究细粒棘球蚴(Echinococcus granulasus)种间遗传标记特点,在内蒙古地区采集羊肝脏中的细粒棘球蚴,或通过手术将患者的包虫剥离囊包后采集细粒棘球蚴,提取虫体DNA,扩增线粒体细胞色素氧化酶1(ND1)基因,将其克隆到PGM-T载体后,用PCR技术鉴定阳性菌落,并进行测序.研究结果表明,细粒棘球蚴DNA扩增出的羊株、人株ND1基因序列片段长度为895bp,ND1基因序列稳定保守,无宿主特异性,可作为细粒棘球蚴理想的种间遗传标记.

In order to investigated the inter-specific genetic marker of Echinococcus granulosus,which was used for further study in molecular classification.The genomic DNA of Echinococcusgranulosuswere collected from the infected sheep and people extracted in Inner Mongolia.The mitochondrial cytochrome oxidase subunit 1（ND1）gene was amplified by PCR using universal primers,and then the PCR product was cloned into pGM-T vectors.The insert was sequenced successfully and compared with other cestodes sequences by DNAStar 5.0and MEGA 4.0.Results showed that the PCR product was 895 bp in length.It suggested that the ND1 gene sequences were stably conservative and non-host-specific,which can be used as an ideal species among Echinococcus granulosus genetic markers.