3种常见人体寄生虫NAD1基因的电子克隆及对比分析
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摘要
目的利用电子克隆法对人体寄生虫NAD1基因进行电子克隆和生物信息学分析,为NAD1基因的进一步深入研究奠定基础。方法分别选取华支睾吸虫、蛔虫和日本血吸虫的一个表达序列标签(EST)片段作为种子序列,以NCBI中的EST数据库作为目标参考序列,用电子克隆的方法分别克隆3种寄生虫的NAD1基因,并对其编码蛋白的理化性质、氨基酸组成、亚细胞定位、二级和三级结构进行对比分析。结果 3种人体寄生虫NAD1蛋白理化性质、亚细胞定位、相对分子量、等电点差异较小,蛔虫和日本血吸虫的NAD1蛋白在三级结构上高度相似。进化分析显示,3种人体寄生虫的NAD1蛋白分属不同分支,存在一定的遗传距离。结论通过电子克隆得到的3个NAD1基因是属于不同种属的同一基因,可广泛应用于研究寄生虫的种间和种内遗传变异。
Objective To in silico clone the NAD1 gene of three common parasites and analyze their bioinformatics,so as to lay the foundation for further research on the NAD gene.Methods By using the in silico cloning method,the full length c DNA(s)of NAD 1 genes of Clonorchis sinensis,Ascaris lumbricoides and Schistosoma japonicum were got,then their physical and chemical properties,compositions of amino acids,subcellular localizations,binary and ternary structures were contrastively analyzed.Results The three kinds of NAD1 proteins were similar in the relative molecular weight,subcellular localization,and physical and chemical properties. The NAD1 proteins were highly similar in binary and ternary structures of A. lumbricoides and S. japonicum. The phylogenetic analysis showed that C. sinensis,A. lumbricoides and S. japonicum belonged to the different evolutionary branches with a certain of genetic distance.Conclusion The three NAD1 genes got from C. sinensis,A. lumbricoides and S. japonicum by in silico cloning belong to the same gene of different species,which can be widely used in the researches of heritable variation of parasites.
Objective To in silico clone the NAD1 gene of three common parasites and analyze their bioinformatics,so as to lay the foundation for further research on the NAD gene.Methods By using the in silico cloning method,the full length c DNA(s)of NAD 1 genes of Clonorchis sinensis,Ascaris lumbricoides and Schistosoma japonicum were got,then their physical and chemical properties,compositions of amino acids,subcellular localizations,binary and ternary structures were contrastively analyzed.Results The three kinds of NAD1 proteins were similar in the relative molecular weight,subcellular localization,and physical and chemical properties. The NAD1 proteins were highly similar in binary and ternary structures of A. lumbricoides and S. japonicum. The phylogenetic analysis showed that C. sinensis,A. lumbricoides and S. japonicum belonged to the different evolutionary branches with a certain of genetic distance.Conclusion The three NAD1 genes got from C. sinensis,A. lumbricoides and S. japonicum by in silico cloning belong to the same gene of different species,which can be widely used in the researches of heritable variation of parasites.
引文
[1]Gill RW,Sanseau P.Rapid in silico cloning of genes using expressed sequence tags(ESTs)[J].Biotechnol Annu Rev,2000,5:25-44.
[2]孙淼,赵茂林.利用表达序列标签电子克隆c DNA全序列的策略[J].生物技术通报,2010,26(1):49-52.
[3]马小娅,庞春英,梁莎莎,等.水牛FADS2基因的电子克隆及序列分析[J].中国畜牧兽医,2017,45(10):2829-2836.
[4]于浩,刘娣,杨秀芹.基于表达序列标签数据库(db EST)的电子克隆技术[J].黑龙江畜牧兽医,2003,46(4):19.
[5]邵筱,吴忠道,刘翰腾,等.应用EST和电子克隆策略研究血吸虫表达基因谱[J].基础医学与临床,2005,25(7):602-606.
[6]Franco GR,Adams MD,Soares MB,et al.Identification of new Schistosoma mansoni genes by the EST strategy using a directional c DNA library[J].Gene,1995,152(2):141-147.
[7]刘媛,蔡嘉斌,蒋国松,等.基于EST的新基因克隆策略[J].遗传,2008,30(3):257-262.
[8]Kennedy CC,Kottom TJ,Limper AH.Characterization of a novel ADAM protease expressed by Pneumocystis carinii[J].Infect Immun,2009,77(8):3328-3336.
[9]Rofatto HK,Tararam CA,Borges WC,et al.Characterization of phosphodiesterase-5 as a surface protein in the tegument of Schistosoma mansoni[J].Mol Biochem Parasitol,2009,166(1):32-41.
[10]张进,刘毅,邹建曾.湖南省鸡蛔虫线粒体nad1基因部分序列的测定及分析[J].中国畜牧兽医,2011,38(2):103-106.
[11]宋慧群,艾琳,莫伊梦,等.片形吸虫线粒体nad1基因的序列分析及种系发育[J].中国兽医科学,2009,39(6):487-491.
[12]Anderson TJ,Blouin MS,Beech RN.Population biology of parasitic nematodes:applications of genetic markers[J].Adv Parasitol,1998,41:219-283.
[13]郑苏月.云南马久邑、洱源日本血吸虫线粒体五个基因片段的研究[D].昆明:昆明医科大学,2014.
[14]何懿菡,孙坤.生物信息学研究进展[J].青海师范大学学报:自科版,2011,27(3):69-72.
[15]王朝辉,马小兵.电子克隆技术及其在医学领域的应用[J].国际生物医学工程杂志,2012,35(6):369-372.
[2]孙淼,赵茂林.利用表达序列标签电子克隆c DNA全序列的策略[J].生物技术通报,2010,26(1):49-52.
[3]马小娅,庞春英,梁莎莎,等.水牛FADS2基因的电子克隆及序列分析[J].中国畜牧兽医,2017,45(10):2829-2836.
[4]于浩,刘娣,杨秀芹.基于表达序列标签数据库(db EST)的电子克隆技术[J].黑龙江畜牧兽医,2003,46(4):19.
[5]邵筱,吴忠道,刘翰腾,等.应用EST和电子克隆策略研究血吸虫表达基因谱[J].基础医学与临床,2005,25(7):602-606.
[6]Franco GR,Adams MD,Soares MB,et al.Identification of new Schistosoma mansoni genes by the EST strategy using a directional c DNA library[J].Gene,1995,152(2):141-147.
[7]刘媛,蔡嘉斌,蒋国松,等.基于EST的新基因克隆策略[J].遗传,2008,30(3):257-262.
[8]Kennedy CC,Kottom TJ,Limper AH.Characterization of a novel ADAM protease expressed by Pneumocystis carinii[J].Infect Immun,2009,77(8):3328-3336.
[9]Rofatto HK,Tararam CA,Borges WC,et al.Characterization of phosphodiesterase-5 as a surface protein in the tegument of Schistosoma mansoni[J].Mol Biochem Parasitol,2009,166(1):32-41.
[10]张进,刘毅,邹建曾.湖南省鸡蛔虫线粒体nad1基因部分序列的测定及分析[J].中国畜牧兽医,2011,38(2):103-106.
[11]宋慧群,艾琳,莫伊梦,等.片形吸虫线粒体nad1基因的序列分析及种系发育[J].中国兽医科学,2009,39(6):487-491.
[12]Anderson TJ,Blouin MS,Beech RN.Population biology of parasitic nematodes:applications of genetic markers[J].Adv Parasitol,1998,41:219-283.
[13]郑苏月.云南马久邑、洱源日本血吸虫线粒体五个基因片段的研究[D].昆明:昆明医科大学,2014.
[14]何懿菡,孙坤.生物信息学研究进展[J].青海师范大学学报:自科版,2011,27(3):69-72.
[15]王朝辉,马小兵.电子克隆技术及其在医学领域的应用[J].国际生物医学工程杂志,2012,35(6):369-372.