白纹伊蚊唾液腺黏蛋白相关蛋白基因1的克
发布时间：2018-09-10 19:57
【摘要】：目的克隆白纹伊蚊唾液腺黏蛋白相关蛋白基因1(Aamucin1)的全长cDNA序列,并分析白纹伊蚊吸血前后基因表达的差异。方法根据白纹伊蚊罗马株Aamucin1基因序列(GenBank序列号为AY826068)设计特异引物,提取白纹伊蚊广州株唾液腺总RNA,PCR扩增Aamucin1全长基因序列,并进行生物信息学分析。实时荧光定量RT-PCR分析白纹伊蚊吸血前后Aamucin1基因表达差异。结果成功克隆白纹伊蚊广州株基因,并获得全长为849 bp的基因序列,编码283个氨基酸。白纹伊蚊广州株与罗马株Aamucin1基因序列比对在C端少了13个氨基酸。与罗马伊蚊同源序列的一致性为58%;该基因具有1个可变剪接子;Protscan分析示有苏氨酸丰富区,可变剪接子恰位于此区;饱血零时(吸血2 h,腹部饱胀者,为BSG_0组),Aamucin1基因表达显著下调,是吸血前(SG组)的0.39倍(P0.01);饱血24 h(饱血后24 h,为BSG_24组),Aamucin1基因表达较饱血零时有回升,是吸血前(SG组)的0.61倍(P0.05)。结论获得白纹伊蚊广州株Aamucin1基因全长序列,并证实其在mRNA水平受到吸血调控。
[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.
【作者单位】： 贵阳医学院寄生虫学教研室;贵阳医学院附属医院;贵阳市第二中学;湖北医药学院;
【基金】：国家自然科学基金(No.30600515,81060138) 贵州省省长基金[黔省专合字(2007)48号]~~
【分类号】：R384
[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.
【作者单位】： 贵阳医学院寄生虫学教研室;贵阳医学院附属医院;贵阳市第二中学;湖北医药学院;
【基金】：国家自然科学基金(No.30600515,81060138) 贵州省省长基金[黔省专合字(2007)48号]~~
【分类号】：R384
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