本文選題：沙蔥螢葉甲　切入點(diǎn)：熱激蛋白　出處：《昆蟲學(xué)報(bào)》2017年08期

【摘要】：【目的】本研究旨在克隆沙蔥螢葉甲Galeruca daurica熱激蛋白Hsp70基因,并對(duì)其進(jìn)行序列和表達(dá)模式分析,探討該基因在沙蔥螢葉甲生長發(fā)育及響應(yīng)溫度脅迫方面的作用�！痉椒ā坎捎肦T-PCR和RACE技術(shù)從沙蔥螢葉甲2齡幼蟲中克隆Hsp70基因,并進(jìn)行生物信息學(xué)分析;用Wo LF PSORT在線軟件進(jìn)行亞細(xì)胞定位預(yù)測;采用實(shí)時(shí)熒光定量PCR檢測該基因在沙蔥螢葉甲成蟲不同組織(頭、胸和腹)中、不同發(fā)育階段(卵、1-3齡幼蟲、蛹、雌雄成蟲)、不同溫度(-14,-10,-5,0,5,10,15,20,25和30℃)處理1 h的2齡幼蟲及0℃下分別處理0 min,15 min,30min,1 h,1.5 h,2 h和3 h時(shí)卵中的相對(duì)表達(dá)量�！窘Y(jié)果】克隆獲得一個(gè)沙蔥螢葉甲Hsp70基因并命名為GdHsp70(GenBank登錄號(hào):KY460462),該基因全長2 340 bp,開放閱讀框(ORF)1 899 bp,編碼632個(gè)氨基酸,預(yù)測蛋白質(zhì)分子量為70.12 k D,等電點(diǎn)(p I)為4.79,無跨膜區(qū),無信號(hào)肽。蛋白質(zhì)亞細(xì)胞定位預(yù)測該蛋白主要位于細(xì)胞質(zhì)內(nèi)。蛋白質(zhì)結(jié)構(gòu)域分析表明,GdHsp70有3個(gè)功能保守區(qū)。同源比對(duì)與系統(tǒng)進(jìn)化分析表明,GdHsp70與分類學(xué)關(guān)系上較為接近的昆蟲的同源蛋白間有較高的相似度。組織特異性和不同發(fā)育階段表達(dá)分析表明,GdHsp70在沙蔥螢葉甲成蟲胸部和卵期表達(dá)量最高;高溫和低溫脅迫均能誘導(dǎo)沙蔥螢葉甲2齡幼蟲體內(nèi)GdHsp70的表達(dá),其中-10℃處理1 h表達(dá)量最高;0℃低溫處理卵15 min至3 h后均能誘導(dǎo)GdHsp70不同程度的表達(dá)上調(diào),其中處理1 h上調(diào)幅度最大�！窘Y(jié)論】沙蔥螢葉甲GdHsp70與該蟲的生長發(fā)育相關(guān),并對(duì)高低溫脅迫的響應(yīng)有重要作用。
[Abstract]:[objective] to clone Galeruca daurica heat shock protein Hsp70 gene and analyze its expression pattern.To investigate the role of this gene in the growth and development of leaf beetle and its response to temperature stress. [methods] the Hsp70 gene was cloned from the 2nd instar larvae by RT-PCR and RACE techniques, and the bioinformatics analysis was carried out.The subcellular localization was predicted by Wo LF PSORT online software, and the gene was detected by real-time fluorescence quantitative PCR in different tissues (head, chest and abdomen) of leaf beetle beetle (head, chest and abdomen).闆岄泟鎴愯櫕),涓嶅悓娓╁害(-14,-10,-5,0,5,10,15,20,25鍜，

本文編號(hào)：1713100