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水稻干尖线虫氨基肽酶基因克隆及功能初步分析

DOI: 10.7668/hbnxb.2015.04.001, PP. 1-7

Keywords: 水稻干尖线虫,氨基肽酶基因,基因克隆,生物信息学分析,原位杂交

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Abstract:

为防治水稻干尖线虫病提供理论支持,研究了水稻干尖线虫氨基肽酶基因特征,并对其编码蛋白结构与功能进行分析。基于水稻干尖线虫转录组测序,并通过RACE技术,从水稻干尖线虫中克隆得到了一氨基肽酶基因Ab-Ampep。生物信息学进行基因分析,原位杂交实验进行表达定位分析。该基因CDS序列全长为2787bp,含有2个开放阅读框(ORF)。最长开放阅读框(ORF)大小2661bp,可编码886个氨基酸,蛋白分子量为101.74kDa,理论等电点(PI)为5.78。编码蛋白氨基酸序列中具有2个Zn2+结合位点和1个PepN保守结构域,有特征信号序列TISHELAHFW,属于谷氨酸锌蛋白(GluZincin)超家族。Ab-Ampep氨基酸序列同挑选的其他9种动物的氨基肽酶氨基酸序列进行多重序列分析,发现功能区域的氨基酸序列较为保守,Ab-Ampep与猪蛔虫(Ascarissuum)的氨基肽酶P(ERG83308.1)蛋白相似度最高为50%。系统进化树分析表明,Ab-Ampep与猪蛔虫(A.suum)的氨基肽酶P(ERG83308.1)蛋白在同一分支上,亲缘关系最近。蛋白质结构分析显示,α-螺旋为Ab-Ampep主要结构元件,能够形成金属离子Zn2+结合位点。原位杂交实验显示,经反义链探针处理的线虫在其生殖部位有颜色加深的杂交信号,该基因的表达位置主要在水稻干尖线虫的生殖部位。通过对水稻干尖线虫氨基肽酶基因研究,利用分子技术改造氨基肽酶或寻找氨基肽酶抑制剂,抑制氨基肽酶的表达,影响线虫生长发育,为防治水稻干尖线虫病提供新思路。

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