本文選題：飛蝗 + 表皮　； 參考：《昆蟲學(xué)報(bào)》2017年03期

【摘要】：【目的】miRNAs(microRNAs)是一類廣泛存在于真核生物中并參與調(diào)控生物體多種生命活動(dòng)的非編碼RNA。昆蟲蛻皮發(fā)育過程包括新表皮的生成和舊表皮的降解。本研究旨在鑒定靶向調(diào)控飛蝗表皮代謝關(guān)鍵基因的miRNAs,為研究飛蝗Locusta migratoria表皮發(fā)育的分子調(diào)控機(jī)制提供一定的實(shí)驗(yàn)基礎(chǔ),同時(shí)為研發(fā)新的害蟲防治分子靶標(biāo)和防治策略提供科學(xué)依據(jù)。【方法】采用生物信息學(xué)方法預(yù)測(cè)與飛蝗表皮代謝關(guān)鍵基因潛在結(jié)合的miRNAs;熒光定量PCR方法檢測(cè)表皮代謝相關(guān)基因及以其為靶標(biāo)的miRNAs在飛蝗2齡和3齡第1,3和5天若蟲中的表達(dá)趨勢(shì);利用免疫共沉淀技術(shù)及雙熒光素酶報(bào)告技術(shù)在體內(nèi)外水平分析miRNA與其靶基因的結(jié)合情況�！窘Y(jié)果】生物信息學(xué)方法預(yù)測(cè)到與脂肪酸合成酶(fatty acid synthase,FAS)、UDP-N-乙酰氨基葡萄糖焦磷酸化酶(UDP-N-acetylglucosamine pyrophorylase,UAP)、糖基轉(zhuǎn)移酶(asparagine-linked glycosylation protein 5,ALG5)和Sinuous等表皮代謝相關(guān)酶基因有潛在結(jié)合能力的miRNAs分別為miRNA-276b,miRNA-2796,miRNA-275和miRNA-184。熒光定量PCR分析表明,FAS,UAP,ALG5以及Sinuous在飛蝗2齡和3齡不同日齡若蟲表皮中具有相似的表達(dá)趨勢(shì),FAS與以其為靶標(biāo)的miRNA-276b表達(dá)趨勢(shì)相同,其余3個(gè)基因與以其為靶標(biāo)的miRNAs的表達(dá)趨勢(shì)相反。通過體內(nèi)免疫共沉淀研究發(fā)現(xiàn),AGO1抗體可顯著富集FAS,UAP,ALG5和Sinuous基因以及以其為靶標(biāo)的miRNAs。體外雙熒光素酶實(shí)驗(yàn)發(fā)現(xiàn),miRNA-276b,miRNA-2796,miRNA-275和miRNA-184對(duì)表皮代謝基因FAS,UAP,ALG5和Sinuous的表達(dá)有較明顯的抑制作用�！窘Y(jié)論】本研究鑒定了靶向調(diào)控飛蝗表皮代謝基因FAS,UAP,ALG5和Sinuous的潛在miRNAs,為進(jìn)一步研究表皮miRNAs對(duì)飛蝗蛻皮發(fā)育的調(diào)控機(jī)制及害蟲防治新靶標(biāo)的發(fā)現(xiàn)提供了重要科學(xué)依據(jù)。
[Abstract]:[objective] miRNAsmicroRNAs) is a class of non-coding RNAs widely found in eukaryotes and involved in the regulation of multiple life activities of organisms. The development of insect molting includes the formation of new epidermis and the degradation of old epidermis. The purpose of this study was to identify the key genes of epidermal metabolism of migratory locust (MiRN), and to provide an experimental basis for studying the molecular regulation mechanism of epidermal development of migratory locust (Locusta migratoria). It also provides scientific basis for the development of new molecular targets and control strategies for pest control. [methods] Bioinformatics was used to predict the potential binding of key genes to epidermis metabolism of locust migratory locust. Fluorescence quantitative PCR method was used to detect epidermis. The expression trend of metabolism-related genes and miRNAs in the 1st and 5th day nymphs of migratory locust at the 2nd and 3rd instar. Immunoprecipitation and double luciferase reporting techniques were used to analyze the binding of miRNA to its target gene in vitro and in vivo. [results] Bioinformatics was used to predict UDP-N- acetyl glucosamine binding to fatty acid synthase. The miRNAs with potential binding ability of UDP-N-acetylglucosamine pyrophorylase, asparagine-linked glycosylation protein 5 (ALG5) and Sinuous genes were miRNA-276bnmiRNA-2796 miRNA-275 and miRNA-184, respectively. Fluorescence quantitative PCR analysis showed that FASA UAPG5 and Sinuous had similar expression trend in the epidermis of second and third instar nymphs of migratory locusts. The expression trend of miRNA-276b was the same as that of miRNA-276b targeting it. The other three genes were opposite to those of miRNAs. The results of immunoprecipitation in vivo showed that the AGO1 antibody could significantly enrich the ALG5 and Sinuous genes of FASA UAPN and its targeting miRNAs. In vitro double luciferase assay showed that miRNA-276bsimiRNA-2796 miRNA-275 and miRNA-184 could significantly inhibit the expression of epidermal metabolite gene FAS-UAPG5 and Sinuous. [conclusion] this study has identified the potential miRNAss that target regulate the epidermal metabolism gene FASUAPALG5 and Sinuous of locust migratory locust. To study the regulation mechanism of epidermal miRNAs on the development of migration locust molt and the discovery of new target for pest control provided important scientific basis.
【作者單位】： 山西大學(xué)應(yīng)用生物學(xué)研究所;山西大學(xué)生命科學(xué)學(xué)院;中國(guó)科學(xué)院動(dòng)物研究所;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31472051,31672364) 山西省科技基礎(chǔ)條件平臺(tái)建設(shè)項(xiàng)目(2015091010)
【分類號(hào)】：S433.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 陳潔;陳宏鑫;姚瓊;張文慶;;甜菜夜蛾UAP的克隆、時(shí)空表達(dá)及RNAi研究[J];中國(guó)農(nóng)業(yè)科學(xué);2014年07期

【共引文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王艷麗;楊美玲;馬恩波;張建珍;;靶向調(diào)控飛蝗表皮代謝相關(guān)基因的miRNA的鑒定與分析[J];昆蟲學(xué)報(bào);2017年03期

【相似文獻(xiàn)】

相關(guān)期刊論文 前8條

1 陳布圣;棉花器官的形態(tài)建成及其生理[J];湖北農(nóng)業(yè)科學(xué);1982年12期

2 霍正浩,樊景禹;大鼠表皮細(xì)胞的體外培養(yǎng)及純化方法[J];動(dòng)物學(xué)雜志;2001年02期

3 秦粉菊,昝林森;黃牛表皮細(xì)胞分離與體外培養(yǎng)最適條件的探索[J];動(dòng)物學(xué)雜志;2003年06期

4 王卅;張e，

本文編號(hào)：1893832