鼠隐藏管状线虫烯醇化酶基因的克隆与序列分析
发布时间：2018-08-30 16:22
【摘要】：为获得鼠隐藏管状线虫(Syphacia obvelata)烯醇化酶结构及其功能,采用同源克隆方法结合RACE技术克隆获得了烯醇化酶基因c DNA序列,并对其进行了序列分析。测序结果显示,扩增获得的鼠隐藏管状线虫烯醇化酶基因大小序列为1603 bp,包含全长为1311 bp的开放阅读框(open reading frame,ORF)序列,共编码436个氨基酸,推导分子量为47.428 k Da。序列分析结果显示,鼠隐藏管状线虫烯醇化酶含有10个丝氨酸激酶磷酸化位点、3个苏氨酸激酶磷酸化位点和4个酪氨酸激酶磷酸化位点,1个潜在的跨膜螺旋结构,无信号肽剪切位点;在亚细胞水平,预测其主要定位于胞浆;二级结构主要以α-螺旋和无规则卷曲为主,其中α-螺旋占34.86%,无规则卷曲占30.50%,延伸链占24.08%,β-转角占10.55%。Swiss-Model模建的3D结构显示,鼠隐藏管状线虫烯醇化酶为一"哑铃"样结构,包含氨基端和羧基端两个结构域,结构域之间为Linker区。每个结构域由多个α-螺旋、β-折叠和卷曲所构成桶形结构,催化中心和Mg~(2+)结合位点位于桶形结构的中心。本研究结果为鼠隐藏管状线虫烯醇化酶基因的功能研究奠定基础。
[Abstract]:In order to obtain the structure and function of (Syphacia obvelata) enolase from hidden tubular nematode of mouse, the c DNA sequence of enolase gene was cloned by homologous cloning method and RACE technique, and the sequence analysis was carried out. The result of sequencing showed that the amplified gene size of rat hidden tubular nematode enolase gene was 1603 bp, containing an open reading frame (open reading frame,ORF (open reading frame,ORF) sequence with a total length of 1311 bp, encoding 436 amino acids and deriving a molecular weight of 47.428 k Da.. Sequence analysis showed that the hidden tubular nematode enolase contained 10 serine kinase phosphorylation sites, 3 threonine kinase phosphorylation sites and 4 tyrosine kinase phosphorylation sites, and a potential transmembrane helical structure. At the subcellular level, it was predicted that it was mainly located in the cytoplasm, and the secondary structure was mainly 伪 -helix and irregular curl. Among them, 伪 -helix was 34.86, irregular curl was 30.50, extended chain was 24.08 and 尾 -angle was used to construct 3D structure of 10.55%.Swiss-Model model. It was found that the hidden tubular nematode enolase was a "dumbbell" like structure, which contained two domains, amino terminal and carboxyl terminal, between which there was a Linker domain. Each domain consists of several 伪 -helix, 尾 -fold and curl structures, and the catalytic center and the Mg~ (2) binding site are located at the center of the bucket structure. The results of this study laid a foundation for the study of the function of the hidden tubular nematode enolase gene in mice.
【作者单位】： 深圳市药品检验研究院;华南农业大学兽医学院;中国农业科学院兰州兽医研究所;深圳市瑞鹏宠物医院;
【基金】：深圳市科技研发资金知识创新计划项目(JCYJ20130402144215888)
【分类号】：Q78;S852.7
本文编号：2213619
[Abstract]:In order to obtain the structure and function of (Syphacia obvelata) enolase from hidden tubular nematode of mouse, the c DNA sequence of enolase gene was cloned by homologous cloning method and RACE technique, and the sequence analysis was carried out. The result of sequencing showed that the amplified gene size of rat hidden tubular nematode enolase gene was 1603 bp, containing an open reading frame (open reading frame,ORF (open reading frame,ORF) sequence with a total length of 1311 bp, encoding 436 amino acids and deriving a molecular weight of 47.428 k Da.. Sequence analysis showed that the hidden tubular nematode enolase contained 10 serine kinase phosphorylation sites, 3 threonine kinase phosphorylation sites and 4 tyrosine kinase phosphorylation sites, and a potential transmembrane helical structure. At the subcellular level, it was predicted that it was mainly located in the cytoplasm, and the secondary structure was mainly 伪 -helix and irregular curl. Among them, 伪 -helix was 34.86, irregular curl was 30.50, extended chain was 24.08 and 尾 -angle was used to construct 3D structure of 10.55%.Swiss-Model model. It was found that the hidden tubular nematode enolase was a "dumbbell" like structure, which contained two domains, amino terminal and carboxyl terminal, between which there was a Linker domain. Each domain consists of several 伪 -helix, 尾 -fold and curl structures, and the catalytic center and the Mg~ (2) binding site are located at the center of the bucket structure. The results of this study laid a foundation for the study of the function of the hidden tubular nematode enolase gene in mice.
【作者单位】： 深圳市药品检验研究院;华南农业大学兽医学院;中国农业科学院兰州兽医研究所;深圳市瑞鹏宠物医院;
【基金】：深圳市科技研发资金知识创新计划项目(JCYJ20130402144215888)
【分类号】：Q78;S852.7
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