高亲和力玉米转磷蛋白基因B103-Zm5473电子克隆及其生物信息学分析

Cloning and bioinformatics analysis of a high affinity phosphate transporter gene B103-Zm5473 in maize

采用电子克隆技术,从玉米自交系B103中克隆出1个高亲和力转磷蛋白基因B103-Zm5473。测序及生物信息学分析结果表明:从B103中克隆的目标DNA序列长度为2 218bp,包含1 608bp构成的开放阅读框(ORF),编码由535个氨基酸组成的蛋白;与玉米标准基因组上注释的基因GRMZM2G045473相比,在DNA序列上存在1个碱基差异,但所编码的蛋白完全相同。生物信息学分析结果表明:来自B103的目的基因编码的B103-Zm5473P蛋白含有转运蛋白FMS超基因家族的高亲和力转磷蛋白Pht1家族蛋白的特征序列GGDYPLSxxIxSE,具有由12个跨膜区和13个亲水区;在13亲水区中,包括N-末端亲水区和C-末端亲水区在内的7个亲水区伸入细胞内部,另外6个亲水区伸出细胞的外部,在第6和第7跨膜区之间存在1个大的亲水区的特征。故该研究中克隆的基因B103-Zm5473,初步推断为Pht1家族的高亲和力转磷蛋白基因。

Here,a high affinity phosphate transporter gene B103Zm5473from the maize inbred line B103was cloned and its bio information was analyzed,with In Silico cloning technique.The results showed that the length of target DNA was2218bp,including functional Open Reading Frame(ORF)encoding a protein comprising of535amino acids in agreed as the annotated gene GRMZM2G045473in the reference maize genome RefGen_B73.The results of bioinformatics analysis revealed that the target protein B103Zm5473P encoded by the gene cloned here had the specific amino acid sequence GGDYPLSxxIxSE of the high affinity phosphate transporter protein of Pht1gene family belonging to FMS super gene family with12transmembrane domains and13hydrophilic domains.Among the13hydrophilic domains,7domains including N and C terminal hydrophilic domains sticked into cell,the other6domains projected over cell,and a huge hydrophilic domain was located between the6th and7th transmembrane domains.In a word,the gene B103Zm5473cloned in present study was to be inferred as encoding a high affinity phosphate transporter protein belonging to Pht1family.