青稞CHI基因的克隆及原核表达分析

Cloning and Prokaryotic Expression Analysis of CHI Gene in Hulless Barley

为了进一步了解CHI基因在青稞黄酮合成中所起的作用,以青稞品系94-19-1为材料,通过同源克隆技术分离CHI基因,并对分离得到的CHI基因进行生物信息学分析及原核表达。结果表明,从青稞94-19-1中克隆得到的CHI基因编码区长为696bp,编码231个氨基酸。序列分析表明,青稞CHI基因有4个碱基和大麦不同,导致1个氨基酸发生改变。SDS-PAGE检测结果表明,将该基因克隆到表达载体pET-32a上,并转化大肠杆菌BL21后,经诱导可成功表达出融合蛋白。

In order to study the role of CHI gene in flavanoids biosynthesis of hulless barley,CHI gene was obtained from hulless barley material＂94-19-1＂though homologous cloning,and the isolated CHI gene was studied by bioinformatics analysis and prokaryotic expression.The results showed a complete open reading frame of 696 bp which encoded 231 amino acids in CHIgene was obtained.By comparing of CHI sequence from both barley and hulless barley,4SNPs were found,which only caused one corresponding amino acid change.The results of SDS-PAGE showed that the fusion protein was successfully expressed by reasonable induction after the linkage of this gene and the expression vector of pET32 awhich transformed into the host bacteriaE.coli BL21.