毛竹β-1,3-葡聚糖酶基因的克隆及序列分析(英文)

β-1,3-葡聚糖酶是一种植物病程相关蛋白,在植物抵御病害中有重要作用。本研究以毛竹为实验材料,利用RACE技术,克隆毛竹β-1,3-葡聚糖酶基因的外显子序列,并在β-1,3-葡聚糖酶基因的编码区两端设计引物,以毛竹基因组DNA为模版扩增该基因的内含子序列。序列结果表明,该基因全长1693bp,包含两个外显子和一个内含子,命名为PheGLU(GenBank登录号GU238236)。该基因包含1个996bp的开放阅读框,编码332个氨基酸,相对分子质量为3.541×104Da,其等电点pI为6.389,是一个酸性蛋白且分泌到胞外;其中,该蛋白的二级结构中包含35.15%的α-螺旋,21.82%的β-转角,43.03%的延伸链和无规则卷曲等;三级结构同源建模预测显示,它与大麦β-1,3-葡聚糖酶(PDB number:1ghsA)具有80.4%的同源性;聚类分析显示,该基因序列与已报道的其它植物具有较的高氨基酸序列同源性。本研究为进一步鉴定毛竹β-1,3-葡聚糖酶基因的抗真菌病害能力奠定了基础。

Cloning and Sequencing Analysis of -1,3-glucanase Gene from Moso Bamboo

β-1,3-glucanase is one kind of plant pathogenesis-related proteins,and plays an important role in plant pathogen defence.In this research,we employed mosot bamboo(Phyllostachys edulis)to be as material for cloning the exon sequence of β-1,3-glucanase gene by using RACE technique,and amplified the intron fragment of B-1,3-glucanase gene from moso bamboo genomic DNA by using primers designed based on its coded region.Sequence analysis showed that the full-length sequence of β-1,3-glucanase gene was 1693 bp,containing two exons and one intron,which named as Phe GLU(GenBank Accession No.GU238236),as well as a ORF(open reading frame)encoded 332 amino acids according to 1 693 bp,which had calculated molecular weight of 3.541 x104Da and predicted isoelectric point of 6.389,it belonged to acidic protein and secreted into the extra cellular space.The secondary structure of this protein harbored 35.15%helix,2 1.82%sheets and 43.03%turns,strands,random coils.etc;its crystal structure similarity to barley β-1,3-glucanase(PDB number:1 ghsA)was as high as 80.4%.The clustering analysis indicated the high amino acids alignment homologies of β-1,3-glucanase with other monocot species.This study would lay a basis for identifying the antifungal ability of β-1,3-glucanase gene.