马铃薯PGA1基因的克隆及序列分析  

Cloning and Sequencing Analysis of PGA1 Gene from Potato

作　　者：闫建俊[1] 白云凤[1] 左静静[1] 裴成成[1] 左敏[1] YAN Jianjun;BAI Yunfeng;ZUO Jingjing;PEI Chengcheng;ZUO Min(Institute of Crop Sciences,Shanxi Academy of Agricultural Sciences,Taiyuan 030031,China)

机构地区：[1]山西省农业科学院作物科学研究所

出　　处：《山西农业科学》2019年第12期2073-2077,共5页Journal of Shanxi Agricultural Sciences

基　　金：山西省青年科技研究基金项目(201701D221210);山西省农业科学院特色农业技术攻关项目(YGG17127);山西省农业科学院作物科学研究所青年基金项目(ZZQ1702)

摘　　要：糖苷生物碱是马铃薯块茎中普遍存在的一种有苦味的毒性甾类生物碱,其毒性可使人、动物变畸形或死亡,PGA1基因是催化糖苷生物碱合成的关键酶基因。通过同源克隆的方法,采用RT-PCR技术对马铃薯大西洋品种的PGA1基因进行克隆,获得了PGA1基因的cDNA序列;并对马铃薯PGA1基因进行生物信息学分析。结果表明,PGA1基因的CDS全长为1497 bp,编码498个氨基酸,所编码蛋白的分子质量为57.087 ku,理论等电点为9.29,为不稳定蛋白;该蛋白的二级结构包括α螺旋(52.41%)、β转角(6.02%)、延伸链(12.85%)和无规则卷曲(28.71%),其中,α螺旋和无规则卷曲为主要元件,与预测的蛋白质三级结构一致。研究结果可为进一步深入了解PGA1基因的功能和调控机制,利用基因工程技术调控马铃薯中糖苷生物碱含量提供理论依据。Steroidal glycoalkaloids taste bitter and poisonous were naturally presented in potato tuber,glycoalkaloids can cause human and animals developmental aberrations or death.PGA1 gene is a key enzyme gene catalyzing the synthesis of glycoside alkaloids.The PGA1 gene of potato was cloned by RT-PCR and the cDNA sequence of PGA1 gene was successfully cloned.Bioinformatics analysis results of PGA1 gene showed that the gene CDS was 1497 bp in length and encoded a protein of 498 amino acids with a predicted molecular quality of 57.087 ku,and theoretical isoelectric point was 9.29.It was an unstable protein.The secondary structure of the potato PGA1 protein included alpha helix(52.41%),βturn(6.02%),extended chain(12.85%)and random coil(28.71%),alpha helix and random coil were the main elements,which were consistent with the predicted protein tertiary structure.This study result will lay a foundation for further understanding of the function and regulatory mechanisms of PGA1 gene and will provide a theoretical basis for regulating the content of glycoside alkaloids in potato by using relevant genetic engineering techniques.

关 键 词：马铃薯 PGA1基因 基因克隆 序列分析 

分 类 号：S532[农业科学—作物学]