Cloning and expression in Escherichia coli of sucrose phosphorylase gene from Leuconostoc mesenteroides No. 165.
Cloning and expression in Escherichia coli of sucrose phosphorylase gene from Leuconostoc mesenteroides No. 165.
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DOI：
10.1271/bbb.60.322
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发表时间：
1996-02
期刊：
影响因子：
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通讯作者：
Haruhiko Kawasaki;Narutoshi Nakamura;Masaaki Ohmori;Takuo Sakai
中科院分区：
文献类型：
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作者：
Haruhiko Kawasaki;Narutoshi Nakamura;Masaaki Ohmori;Takuo Sakai
采用PCR方法扩增了一株大肠杆菌（Leuconostoc mesenteroides No. 165）的蔗糖磷酸化酶基因，克隆于pUC118上，并在大肠杆菌中表达。该基因的核苷酸序列与肠系膜乳杆菌ATCC12291相似度为96.3%，与变形链球菌相似度为67%，与葡萄农杆菌基因相似度较低。克隆的基因与lacZalpha融合，在大肠杆菌中与IPTG诱导表达，产生了大量的活性酶，约占细胞总蛋白的50%。
The sucrose phosphorylase gene of an isolate, Leuconostoc mesenteroides No. 165, was amplified by PCR, cloned on pUC118, and expressed in E. coli. The nucleotide sequence of the gene showed 96.3% similarity to that of L. mesenteroides ATCC12291 and 67% to that of Streptococcus mutans, but low similarity to the Agrobacterium vitis gene. The cloned gene, which fusing with lacZalpha, was expressed inducibly with IPTG in E. coli to produce an active enzyme in large quantities that accounted for about 50% of the total cell protein.