菠菜乙醇酸氧化酶在大肠杆菌中的克隆与表达
[Cloning and expression of spinach glycolate oxidase in Escherichia coli].
作者信息
Jin Jian-Feng, Tan Tian-Wei, Su Guo-Fu
机构信息
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
出版信息
Sheng Wu Gong Cheng Xue Bao. 2002 Jan;18(2):212-5.
The cDNA coding spinach glycolate oxidase (GO) was amplified by RT-PCR using the total RNA of spinach leaves as the template, and was cloned into cloning vector pMD18-T. After the DNA sequence was determined, the go gene was subcloned into E. coli expression vector pBV220, pET-22b(+), pTIG-Trx and pThioHisC. SDS-PAGE analysis revealed that the recombinant E. coli BL21 (DE3) (pTIG-Trx-GO) and E. coli BL21 (DE3) (pET-22b(+)-GO) expressed the predicted 38 kD glycolate oxidase, and the enzyme activity was also detected.
以菠菜叶片总RNA为模板，通过RT-PCR扩增编码菠菜乙醇酸氧化酶（GO）的cDNA，并将其克隆到克隆载体pMD18-T中。测定DNA序列后，将go基因亚克隆到大肠杆菌表达载体pBV220、pET-22b(+)、pTIG-Trx和pThioHisC中。SDS-PAGE分析表明，重组大肠杆菌BL21(DE3)(pTIG-Trx-GO)和大肠杆菌BL21(DE3)(pET-22b(+)-GO)表达了预测的38 kD乙醇酸氧化酶，并且还检测到了酶活性。
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