大豆乙醇酸氧化酶基因的克隆、表达和酶学活性分析

摘要

The cDNA sequence of glycolate oxidase (GO) was amplified from the total RNA of soybean leaves by RT-PCR,and then cloned into cloning vector pMD19-T simple. After sequencing, the GO gene was transformed into Escherichia coli expression vector pRSET-a. SDS-PAGE anaksis proved that the recombinant E. coli BL21 expressed the predicted 40.8kD glycolate oxidase; and the enzyme activity was also detected.%利用RT-PCR技术,从大豆叶片总RNA中扩增了乙醇酸氧化酶(GO)基因的cDNA序列,克隆到pMD19-T simple上,进行测序,然后将乙醇酸氧化酶的cDNA克隆至原核表达载体pRSET-A上,转化E.coli BL21,并对该基因在 E.coli中的表达进行了研究.