利用mRNA差异显示技术分离捻转血矛线虫感染性三期幼虫抗干燥相关基因  

Isolation of desiccation survival related genes from infective third-stage larvae of Haemonchus contortus by mRNA differential display technique

作　　者：陈学秋[1] 马羽洁[1] 周前进[1] 张红丽[1] 杨怡[1] 闫宝龙[1] 杜爱芳[1] 

机构地区：[1]浙江大学动物科学学院,浙江杭州310058

出　　处：《中国兽医学报》2013年第8期1212-1216,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资金资助项目(No.2011XZZX006)

摘　　要：捻转血矛线虫感染性三期幼虫对干燥环境具有很强的抵抗力,环境适宜时可恢复到正常状态并继续感染宿主。为初步探讨捻转血矛线虫感染性三期幼虫抗干燥的分子机制,试验对该期幼虫施以不同干燥条件的处理,筛选出虫体存活率达到90%的干燥条件。在筛选到的条件下(相对湿度为50%,时间为60h)处理虫体,采用mRNA差异显示技术分析干燥环境下培养的虫体和正常虫体的mRNA表达差异。经验证获得了48个差异表达的EST序列,其中25个(52.08%)序列与现有捻转血矛线虫基因组数据库有高度同源性;15个(31.25%)序列与已知线虫的cDNA序列或基因组序列有较高同源性;9个(18.75%)序列与其他线虫已知蛋白质氨基酸序列同源性较高。差异表达序列AF-U01C与多种线虫的蛋白酶体β亚基家族成员同源性达90%,差异表达序列AG-U01A与多种线虫的β酮脂酰CoA硫解酶(3-ketoacyl-CoA thiolase)同源性达65%。结果为进一步研究寄生性线虫抗干燥机制奠定基础。Infective third-stage larvae of Haemonchus contortus have great resistance to desiccation, when environment gets favorable the larvae can resume normal life and infect hosts. To understand the molecular mechanism of infective H. contortus L3s larvae activated during anhydrobiosis, a condition that can maintain the RH （relative humidity） as low as possible and ensure the survival rate of nematodes as high as 90% was selected by a preliminary experiment. Under the condition selected （50% RH for 60 h） ,H. contortus L3s were cultured to identify the differentially expressed genes compared to the control nematodes by mRNA differential display technique. 48 transcripts differentially expressed were obtained during desiccation stress, of which 25 （52. 08%） matched publicly available genomic DNA sequences of H. contortus, 15 （31.25%） matched known cDNA se-quences or genomic DNA sequences of nematodes and 9（18.75%） matched known proteins of oth-er nematodes. The sequence AF-U01C had 90% identity with proteasome Beta subunit family member of other nematodes,AG-U01A had 65% identity with long-chain 3-ketoacyl-CoA thiolase. The results provide a funding for further research into the mechanism of desiccation survival in parasitic nematodes.

关 键 词：捻转血矛线虫感染性三期幼虫 干燥相关基因 MRNA差异显示技术 

分 类 号：S852.731[农业科学—基础兽医学]