Cloning and Expression of a Gene for an 87-kDa β-1,3-Glucanase of Bacillus circulans IAM1165 in Escherichia coli K-12
Cloning and Expression of a Gene for an 87-kDa β-1,3-Glucanase of Bacillus circulans IAM1165 in Escherichia coli K-12
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环状芽孢杆菌 IAM1165 87-kDa β-1,3-葡聚糖酶基因在大肠杆菌 K-12 中的克隆和表达
DOI：
10.1271/bbb.56.1854
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
K. Horikoshi
中科院分区：
文献类型：
--
作者：
R. Aono;Mami Yamamoto;Masakazu Sato;Y. Nogi;K. Horikoshi
将来自环状芽孢杆菌IAM 1165（能够裂解真菌细胞壁的β-1，3-葡聚糖酶的高产菌）的染色体DNA的部分Sau 3AI消化片段插入质粒载体pHSG 399的BamHI位点。通过鸟枪法在大肠杆菌K-12中克隆了葡聚糖酶的基因。利用8 kb插入DNA在大肠杆菌中定向合成87 kDa内切-β-1，3-n-葡聚糖酶。杆菌β-葡聚糖酶基因位于插入DNA中的2.6-kb EcoRI-SmaI片段中。酶活性主要存在于E.携带该基因的大肠杆菌。
Partially Sau3AI-digested fragments of chromosomal DNA from Bacillus circulans IAM1165, a high producer of β-1,3-glucanases able to lyse fungal cell walls, were inserted into a BamHI site of the plasmid vector pHSG399. A gene for the glucanase was cloned in Escherichia coli K-12 by the shotgun method. An 8-kb inserted DNA directed synthesis of an 87-kDa endo-β-1,3-n-glucanase in E. coli. The β-glucanase gene was in a 2.6-kb EcoRI-SmaI segment within the insert DNA. The enzyme activity was found mainly in the periplasmic fraction of E. coli carrying the gene.