尼泊尔黄堇G6PDH基因的克隆和生物信息学分析

Cloning and Bioinformatics Analysis of G6PDH Gene in Corydalis hendersonii Hemsl.

为了深入发掘该基因在尼泊尔黄堇生态适应和次生代谢物积累中的作用机制,本研究依据转录组中葡萄糖-6-磷酸脱氢酶(G6PDH)基因的注释信息,应用聚合酶链式反应(PCR)技术克隆了尼泊尔黄堇G6PDH基因,通过生物信息学方法进行相关分析。结果表明:克隆的G6PDH基因的开放阅读框(ORF)为1581 bp,编码526个氨基酸。系统进化树分析表明尼泊尔黄堇G6PDH蛋白氨基酸序列与博落回、罂粟等具有很高的相似度,尤其与同是罂粟科的博落回相似度最高,相似度达到91%。通过一系列生物信息学分析发现,ChG6PDH蛋白分子量为59.83 kD;理论等电点为6.37;该蛋白为不稳定、亲水性、非分泌性蛋白,无跨膜结构域;二级结构中具体α-螺旋(40.11%)和β-折叠(6.08%)、无规则卷曲(40.87%)以及延伸链(12.93%)结构,预测得到的三级结构中具有典型的NADPH结合位点。该基因的成功克隆和预测结果将为进一步研究ChG6PDH基因在尼泊尔黄堇生长发育、逆境适应和次生代谢物积累等方面中的功能提供帮助。

Based on the annotation information of G6 PDH gene in transcriptome,this study cloned G6 PDH gene of Corydalis hendersonii Hemsl.by polymerase chain reaction(PCR),and its correlation was analyzed by bioinformatics method.In order to explore the mechanism of G6 PDH gene in ecological adaptation and secondary metabolite accumulation of Corydalis hendersonii.The results showed that the ORF of G6 PDH gene was 1581 bp,encod ing 526 amino acids.Phylogenetic tree analysis showed that the amino acid sequence of G6 PDH protein of Corydalis hendersonii had a high similarity with Macleaya cordata and Papaver somniferum L.,especially with the same family of Papaver somniferum,the similarity reached 91%.Through a series of bioinformatics analysis,it is found that the molecular weight of ChG6 PDH protein is 59.83 kD;the theoretical isoelectric point is 6.37;the protein is an unstable,hydrophilic,non secretory protein without transmembrane domain;the specificα-helix(40.11%)andβ-fold(6.08%),random coil(40.87%)and extension chain(12.93%)structures in the secondary structure,and there are typical NADPH binding sites in predict tertiary structure.The successful cloning of ChG6 PDH gene and the bioinformatics analysis results of ChG6 PDH protein will be helpful to further study the function of the ChG6 PDH gene in the growth,adaptation to adversity and accumulation of secondary metabolites of Corydalis hendersonii Hemsl.