芸香草CdLEA3基因的克隆及原核表达分析

Cloning and Prokaryotic Expression of CdLEA3 Gene from Cymbopogon distans

本研究以芸香草为试验材料,采用RT-PCR和RACE技术对芸香草CdLEA3基因进行了分离克隆以及原核分析,并进行了生物信息学分析与原核表达分析。结果显示,芸香草CdLEA3基因的cDNA全长为934 bp,含627 bp的最大开放阅读框,编码208个氨基酸;生物信息学分析表明,CdLEA3基因编码蛋白的分子质量预测为21.6 kD,包含75.5%的亲水性氨基酸,不稳定指数为34.5,具有极强的亲水性,属于稳定性强的亲水性蛋白。CdLEA3基因编码蛋白的二级结构主要为α-螺旋结构;SDS-PAGE分析表明CdLEA3蛋白分子质量约为22 kD,且全部在上清液中表达。本研究通过对芸香草CdLEA3基因克隆分析,为进一步研究CdLEA3基因的分子作用机制及功能提供科学依据。

In this study,rutaceous was used as experimental material,RT-PCR and RACE technology were used to isolate and clone the CdLEA3 gene of rutaceous and conduct the prokaryotic analysis,as well as the bioinformatics analysis and prokaryotic expression analysis.The results showed that the full length of the cDNA of the CdLEA3 gene was 934 bp,containing a largest open reading frame of 627 bp,encoding 208 amino acids,and the predicted molecular weight of the protein was 21.6 k D,containing 75.5%hydrophilic amino acid,and the instability index was 34.5.It was highly hydrophilic and belonged to the hydrophilic protein with strong stability.And the secondary structure of the protein encoded by CdLEA3 gene was mainlyα-helix;SDS-PAGE analysis showed that the molecular weight of CdLEA3 protein was about 22 k D,and all of them were expressed in supernatant.The CdLEA3 gene was cloned and analyzed to provide a scientific basis for the further study of the molecular mechanism and function of CdLEA3 gene.