【摘要】：為獲得鼠隱藏管狀線蟲(Syphacia obvelata)烯醇化酶結(jié)構(gòu)及其功能,采用同源克隆方法結(jié)合RACE技術(shù)克隆獲得了烯醇化酶基因c DNA序列,并對(duì)其進(jìn)行了序列分析。測序結(jié)果顯示,擴(kuò)增獲得的鼠隱藏管狀線蟲烯醇化酶基因大小序列為1603 bp,包含全長為1311 bp的開放閱讀框(open reading frame,ORF)序列,共編碼436個(gè)氨基酸,推導(dǎo)分子量為47.428 k Da。序列分析結(jié)果顯示,鼠隱藏管狀線蟲烯醇化酶含有10個(gè)絲氨酸激酶磷酸化位點(diǎn)、3個(gè)蘇氨酸激酶磷酸化位點(diǎn)和4個(gè)酪氨酸激酶磷酸化位點(diǎn),1個(gè)潛在的跨膜螺旋結(jié)構(gòu),無信號(hào)肽剪切位點(diǎn);在亞細(xì)胞水平,預(yù)測其主要定位于胞漿;二級(jí)結(jié)構(gòu)主要以α-螺旋和無規(guī)則卷曲為主,其中α-螺旋占34.86%,無規(guī)則卷曲占30.50%,延伸鏈占24.08%,β-轉(zhuǎn)角占10.55%。Swiss-Model模建的3D結(jié)構(gòu)顯示,鼠隱藏管狀線蟲烯醇化酶為一"啞鈴"樣結(jié)構(gòu),包含氨基端和羧基端兩個(gè)結(jié)構(gòu)域,結(jié)構(gòu)域之間為Linker區(qū)。每個(gè)結(jié)構(gòu)域由多個(gè)α-螺旋、β-折疊和卷曲所構(gòu)成桶形結(jié)構(gòu),催化中心和Mg~(2+)結(jié)合位點(diǎn)位于桶形結(jié)構(gòu)的中心。本研究結(jié)果為鼠隱藏管狀線蟲烯醇化酶基因的功能研究奠定基礎(chǔ)。
[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 偽 -helix and irregular curl. Among them, 偽 -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 偽 -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.
【作者單位】： 深圳市藥品檢驗(yàn)研究院;華南農(nóng)業(yè)大學(xué)獸醫(yī)學(xué)院;中國農(nóng)業(yè)科學(xué)院蘭州獸醫(yī)研究所;深圳市瑞鵬寵物醫(yī)院;
【基金】：深圳市科技研發(fā)資金知識(shí)創(chuàng)新計(jì)劃項(xiàng)目(JCYJ20130402144215888)
【分類號(hào)】：Q78;S852.7

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