捻转血矛线虫感染性三期幼虫gst基因的筛选及抗干燥功能分析  

Screening and analysis of gst gene related to desiccation resistance of Haemonchus contortus infective third-stage larvae

作　　者：马羽洁[1] 陈学秋[1] 闫宝龙[1] 杨怡[1] 郭筱璐 杜爱芳[1] 

机构地区：[1]浙江大学动物预防医学研究所/浙江省动物预防医学重点实验室,浙江杭州310058

出　　处：《中国兽医学报》2015年第3期401-405,共5页Chinese Journal of Veterinary Science

基　　金：高等学校博士学科点专项科研基金资助项目(20100101110102);中央高校基本科研业务费专项基金资助项目(2011XZZX006)

摘　　要：为研究干燥作用对捻转血矛线虫L3期幼虫mRNA表达的影响,筛选并分析干燥相关表达基因,本试验利用mRNA差异显示技术得到了捻转血矛线虫干燥相关基因,通过相对荧光定量PCR验证所获基因的表达情况,并在秀丽隐杆线虫中通过RNA干扰抑制相关同源基因的表达检测干燥后复苏虫体生存率以研究基因抗干燥功能。结果显示,差异表达序列GH-U02A与已知捻转血矛线虫谷胱甘肽-S-转移酶(GST)氨基酸同源性为94%。对秀丽隐杆线虫Ce-gst-7基因进行RNA干扰后,虫体在干燥后的生存率降低,证明gst基因在线虫中有抗干燥功能。With the aim to investigate the differentially expressed genes related to desiccation survival of Haemonchus contortus infective L3 larvae,mRNA differential display technique was used.And the selected candidate genes were assayed by comparative quantitative real-time PCR.Furthermore,the homologous genes in C.elegans were treated with RNAi to verify the function of them on desiccation tolerance.The results revealed that differentially expressed sequence GHU02 Ahas 94% amino acid homology with the known Haemonchus contortus glutathione S-transferase(GST).Supression of Ce-gst-7gene expression by RNAi in C.elegans showed a decline of survival rate during desiccation.The results confirmed that gst gene is resistant to desiccation when nematodes were placed in dry environment.

关 键 词：捻转血矛线虫L3期幼虫 干燥 银染mRNA差异显示技术 GST RNA干扰 

分 类 号：S852.7[农业科学—基础兽医学]