捻转血矛线虫丙硫咪唑耐药株和敏感株转录组水平下差异基因分析  

Differential gene analysis at the transcriptome level of albendazole resistant and sensitive strains of Haemonchus contortus

作　　者：石雅琴 苏倩 王腾宇 毛晓伟 陈昕迪 穆嘉明 王文龙 刘春霞[2] SHI Yaqin;SU Qian;WANG Tengyu;MAO Xiaowei;CHEN Xindi;MU Jia-ming;WANG Wen-long;LIU Chunxia(Ministry of Agriculture Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease,College of Veterinary Medicine,Inner Mongolia Agri-cultural University,Hohhot 010018,China;College of Life Sciences,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学兽医学院农业部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018 [2]内蒙古农业大学生命科学院,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2021年第7期1295-1300,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31760731);内蒙古自治区科技计划资助项目(201702074)。

摘　　要：首先筛选捻转血矛线虫丙硫咪唑敏感株和耐药株单核苷酸多态性(single nucleotide polymorphism,SNP)信息,然后从比较转录组测序数据中筛选出4个丙硫咪唑耐药相关突变基因HCON_00192020、HCON_00037800、HCON_00129090和HCON_00178650,并进行生物信息学分析,最后对差异基因进行PCR扩增并测序。结果显示,共筛选到41个具有SNP位点的基因,选择reads≥10的4个基因进行后续基因扩增和生物信息学分析。扩增后目的条带大小均正确,测序后显示HCON_00192020基因在3个耐药虫株中第266位均由T突变为C,HCON_00037800基因在3个耐药虫株中第179、182、188位均由A突变为G,HCON_00129090基因在3个耐药虫株中第283位由G突变为A,HCON00178650基因在3个耐药虫株中第89位均由C突变为A,突变结果均与转录组测序结果一致。SNP information of albendazole sensitive and resistant strains of Haemonchus contortus was screened,and then four albendazole resistant mutant genes were screened from the comparative transcriptome sequencing data HCON_00192020,HCON_00037800,HCON_00129090 and HCON_00178650.Finally,the differential genes were amplified by PCR and sequenced.The results showed that a total of 41 genes with SNP sites were selected,and 4 genes with reads≥10 were selected for subsequent gene amplification and bioinformatics analysis.Amplification results showed that the size of the target bands was correct.Consistent with transcriptome sequencing results,the analysis of sequencing result of PCR products showed the 266 th nucleotide of HCON_00192020 gene was mutated from T to C in the three drug-resistant insect strains,and the 179 th,182 th,188 th nucleotides of the HCON_00037800 gene of the three drug-resistant insect strains were mutated from A to G.The HCON_00129090 gene was mutated from G to A at position 283 in the three drug-resistant insect strains.The HCON_00178650 gene was mutated from C to A at position 89 of the three drug-resistant insect strains.The above research can be used for the study of drug resistance generation mechanism and clinical drug resistance detection.

关 键 词：捻转血矛线虫 丙硫咪唑 耐药性 单核苷酸多态性 

分 类 号：S852.7[农业科学—基础兽医学]