【摘要】：目的克隆白紋伊蚊唾液腺黏蛋白相關(guān)蛋白基因1(Aamucin1)的全長(zhǎng)cDNA序列,并分析白紋伊蚊吸血前后基因表達(dá)的差異。方法根據(jù)白紋伊蚊羅馬株Aamucin1基因序列(GenBank序列號(hào)為AY826068)設(shè)計(jì)特異引物,提取白紋伊蚊廣州株唾液腺總RNA,PCR擴(kuò)增Aamucin1全長(zhǎng)基因序列,并進(jìn)行生物信息學(xué)分析。實(shí)時(shí)熒光定量RT-PCR分析白紋伊蚊吸血前后Aamucin1基因表達(dá)差異。結(jié)果成功克隆白紋伊蚊廣州株基因,并獲得全長(zhǎng)為849 bp的基因序列,編碼283個(gè)氨基酸。白紋伊蚊廣州株與羅馬株Aamucin1基因序列比對(duì)在C端少了13個(gè)氨基酸。與羅馬伊蚊同源序列的一致性為58%;該基因具有1個(gè)可變剪接子;Protscan分析示有蘇氨酸豐富區(qū),可變剪接子恰位于此區(qū);飽血零時(shí)(吸血2 h,腹部飽脹者,為BSG_0組),Aamucin1基因表達(dá)顯著下調(diào),是吸血前(SG組)的0.39倍(P0.01);飽血24 h(飽血后24 h,為BSG_24組),Aamucin1基因表達(dá)較飽血零時(shí)有回升,是吸血前(SG組)的0.61倍(P0.05)。結(jié)論獲得白紋伊蚊廣州株Aamucin1基因全長(zhǎng)序列,并證實(shí)其在mRNA水平受到吸血調(diào)控。
[Abstract]:Objective to clone the full-length cDNA sequence of salivary gland mucin-associated protein gene 1 (Aamucin1) from Aedes albopictus and analyze the difference of gene expression before and after blood sucking. Methods based on the Aamucin1 gene sequence of Aedes albopictus Roman strain (GenBank sequence number is AY826068), a specific primer was designed to amplify the full-length Aamucin1 gene sequence from the total salivary gland of Guangzhou Aedes albopictus strain, and the sequence was analyzed by bioinformatics. The difference of Aamucin1 gene expression between Aedes albopictus and Aedes albopictus was analyzed by real-time fluorescence quantitative RT-PCR. Results the gene of Guangzhou strain of Aedes albopictus was cloned successfully, and the total length of 849 bp gene was obtained, encoding 283-amino acid. The sequence alignment of Aamucin1 gene between Guangzhou strain and Roman strain of Aedes albopictus lost 13 amino acids at C terminal. The homology of the sequence with Aedes Romanus is 58. The gene has a variable splice region, which is located in this region, and 00:00 full blood (2 h sucking, abdominal fullness), the results of Protscan analysis show that the gene has a threonine rich region, and the variable splice is located in the same region. The expression of Aamucin1 gene in BSG_0 group was significantly down-regulated, 0.39 times as much as that in SG group (P0.01), and the expression of Aamucin1 gene increased significantly at 24 h after satiety (24 h after satiety, in BSG_24 group), and was 0.61 times higher than that before SG (P0.05). Conclusion the full-length sequence of Aamucin1 gene of Aedes albopictus Guangzhou strain was obtained, and it was confirmed that Aedes albopictus was regulated by blood sucking at mRNA level.
【作者單位】： 貴陽(yáng)醫(yī)學(xué)院寄生蟲(chóng)學(xué)教研室;貴陽(yáng)醫(yī)學(xué)院附屬醫(yī)院;貴陽(yáng)市第二中學(xué);湖北醫(yī)藥學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(No.30600515,81060138) 貴州省省長(zhǎng)基金[黔省專(zhuān)合字(2007)48號(hào)]~~
【分類(lèi)號(hào)】：R384

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