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| 链霉菌(Streptomyces ST66)MalQ基因克隆与原核融合表达 | |
| 引用本文： | 王水兴,付金衡,龚珩,郭勇,夏慧玲,韩林强.链霉菌(Streptomyces ST66)MalQ基因克隆与原核融合表达[J].农业生物技术学报,2007,15(6):1024-1028. |
| 作者姓名： | 王水兴 付金衡 龚珩 郭勇 夏慧玲 韩林强 |
| 作者单位： | 1. 南昌大学中德联合研究院,南昌,330047 2. 九江学院医学院,九江,332005 3. 华南理工大学生物科学与工程学院,广州,510641 |
| 摘 要： | 利用PCR方法获得Streptomyces ST66 MalQ基因,将该基因插入原核表达载体pTrc-CKS中,对质粒所含外源片段双向测序,并经BLAST比较分析,发现其与GenBank中报道的Streptomyces coelicolor MalQ基因的同源性高达97%。重组质粒pTrc-MalQ转化大肠杆菌(Escherichia coli)Top10F',经IPTG诱导后以融合蛋白形式表达,SDS-PAGE电泳显示MalQ基因与CKS(CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase or CMP-KDO synthetase)基因表达的融合蛋白在目标位置约106kD处有明显条带。经薄层层析分析粗酶液处理的麦芽三糖溶液,证实粗酶液具有麦芽糖转糖基活性。 |
| 关 键 词： | 麦芽糖转糖基酶 基因克隆 融合表达 大环糊精 MalQ基因 |
| 文章编号： | 1006-1304（2007）06-1024-05 |
| 收稿时间： | 2007-06-18 |
| 修稿时间： | 2007-07-19 |
Cloning and Fusion Expression of the MalQ Gene from Streptomyces ST66 |
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| WANG Shui-xing,FU Jin-heng,GONG Heng,GUO Yong,XIA Hui-ling,HAN Lin-qiang.Cloning and Fusion Expression of the MalQ Gene from Streptomyces ST66[J].Journal of Agricultural Biotechnology,2007,15(6):1024-1028. | |
| Authors: | WANG Shui-xing FU Jin-heng GONG Heng GUO Yong XIA Hui-ling HAN Lin-qiang |
| Abstract: | The MalQ gene was amplified from Streptomyces ST66 by PCR, cloned into pTrc-CKS and sequenced. MalQ sequence was analyzed by BLAST and displayed high similarity(97%) to MalQ gene of Streptomyces coelicolor reported in GenBank. The recombined plasmid pTrc- MalQ was transformed into Escherichia coli Top 10F' and induced by IPTG. MalQ gene and CKS(CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase or CMP-KDO synthetase)gene fusion protein(molecular weight:about 106 kD) could be detected by SDS-PAGE gel electrophoresis. The crude enzyme was demonstrated having 4-glucanotransferase activity with thin layer chromatography. |
| Keywords: | amylomaltase gene clone fusion expression cycloamylose MalQ gene |
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