淡色庫蚊ATP合酶B亞基基因克隆和序列分析及其與溴氰菊酯抗性的關(guān)系
發(fā)布時間:2018-01-20 02:00
本文關(guān)鍵詞: 淡色庫蚊 ATP合酶B亞基 抗藥性 溴氰菊酯 生物信息學(xué) 出處:《昆蟲學(xué)報》2017年08期 論文類型:期刊論文
【摘要】:【目的】克隆淡色庫蚊Culex pipiens pallens ATP合酶B亞基基因編碼區(qū)序列,并進行生物信息學(xué)分析,研究其與溴氰菊酯抗性關(guān)系!痉椒ā客ㄟ^PCR方法擴增ATP合酶B亞基基因編碼區(qū)序列;利用生物信息學(xué)網(wǎng)站在線分析工具,預(yù)測ATP合酶B亞基基因編碼蛋白的理化性質(zhì)和功能特征;通過實時定量PCR方法比較ATP合酶B亞基基因在室內(nèi)篩選的淡色庫蚊溴氰菊酯敏感品系和抗性品系中的表達,并在現(xiàn)場種群溴氰菊酯敏感和抗性個體中做進一步驗證!窘Y(jié)果】成功克隆淡色庫蚊ATP合酶B亞基基因編碼區(qū)序列(Gen Bank登錄號:KY783434),長717 bp,編碼238個氨基酸。生物信息學(xué)分析表明,其編碼蛋白理論相對分子量為26.96 k D,等電點為8.97,在第70-231位氨基酸具有ATP合酶B亞基結(jié)構(gòu)域,未發(fā)現(xiàn)信號肽和跨膜區(qū)。實時定量PCR結(jié)果顯示,ATP合酶B亞基基因在室內(nèi)篩選的淡色庫蚊抗溴氰菊酯品系和現(xiàn)場種群溴氰菊酯抗性個體中表達量均上調(diào)!窘Y(jié)論】本研究獲得了淡色庫蚊ATP合酶B亞基基因編碼區(qū)序列,進行了生物信息學(xué)分析,并證實其在溴氰菊酯抗性個體中高表達,為進一步研究該基因在蚊抗藥性中的作用奠定了基礎(chǔ)。
[Abstract]:[objective] to clone the coding region of subunit B of Culex pipiens pallens ATP synthase from Culex pipiens pallens and analyze it by bioinformatics. [methods] the coding region of ATP synthase B subunit gene was amplified by PCR. The physical and chemical properties and functional characteristics of ATP synthase B subunit gene encoding protein were predicted by online analysis tools of bioinformatics website. The expression of ATP synthase B subunit gene in susceptible and resistant strains of Culex pipiens pallens was compared by real-time quantitative PCR. The results were further verified in the susceptible and resistant individuals of the field population. [results] the coding region of ATP synthase B subunit gene of Culex pipiens pallens was cloned successfully (. Gen Bank: KY783434. The length of 717 BP encodes 238 amino acids. Bioinformatics analysis shows that the theoretical molecular weight of the encoded protein is 26.96 KD and the isoelectric point is 8.97. The ATP synthase B subunit domain was found at the 70-231 amino acid site, and no signal peptide or transmembrane region was found. The results of real-time quantitative PCR showed that there was no signal peptide or transmembrane domain. The expression of ATP synthase B subunit gene was up-regulated in the deltamethrin resistant strains of Culex pipiens pallens and in the field population. [conclusion] the ATP synthase B subunit of Culex pipiens pallens was obtained in this study. Sequence of gene coding region. Bioinformatics analysis was carried out, and its high expression in deltamethrin resistant individuals was confirmed, which laid a foundation for further study on the role of the gene in mosquito resistance.
【作者單位】: 河北醫(yī)科大學(xué)病原生物學(xué)教研室;南京醫(yī)科大學(xué)病原生物學(xué)系;保定市第二中心醫(yī)院血液腎病科;蚌埠市第三人民醫(yī)院檢驗科;
【基金】:河北省高等學(xué)?茖W(xué)技術(shù)研究項目(QN2016005)
【分類號】:R384.1
【正文快照】: 蚊可以傳播瘧疾、淋巴絲蟲病、登革熱、寨卡病毒病等多種蚊媒病,嚴重危害人類健康(Reiner etal.,2013;Mayer et al.,2017)。長期以來,化學(xué)防制在控制蚊媒、預(yù)防蚊媒病中具有重要作用(Scott etal.,2015)。但大量文獻報道,蚊對一種或多種殺蟲劑已經(jīng)產(chǎn)生了不同程度抗性(梁軍等,20
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