Molecular cloning of staphylococcal enterotoxin B gene in Escherichia coli and Staphylococcus aureus.
Molecular cloning of staphylococcal enterotoxin B gene in Escherichia coli and Staphylococcus aureus.
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大肠杆菌和金黄色葡萄球菌中葡萄球菌肠毒素 B 基因的分子克隆。
DOI：
10.1073/pnas.82.17.5850
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发表时间：
1985
影响因子：
11.1
通讯作者：
Khan,SA
中科院分区：
文献类型：
--
作者：
Ranelli,DM;Jones,CL;Johns,MB;Mussey,GJ;Khan,SA
我们以pBR322为载体，在大肠杆菌中克隆了金黄色葡萄球菌entB基因，但该克隆不产生可检测到的葡萄球菌肠毒素B(SEB)。当entB基因被置于强噬菌体启动子PR的下游时，在大肠杆菌中可以很容易地检测到SEB的合成。有趣的是，成熟的SEB几乎只存在于细胞质部分。发现SEB前体与细胞膜相关。将entB基因重新导入金黄色葡萄球菌，得到的克隆能产生SEB。EntB基因被定位在一个2.1kb的区域。对entB基因的一部分进行Maxam-Gilbert测序，得到了与SEB已知氨基酸序列相对应的DNA序列。Southern杂交实验表明，在SEB产生菌的染色体中，entB基因存在于相同的限制性内切酶片段上。在SEB非产生菌中，entB基因缺失。
We have cloned the Staphylococcus aureus entB gene in Escherichia coli, using pBR322 as the vector plasmid; however, no detectable staphylococcal enterotoxin B (SEB) was produced by the E. coli clones. When the entB gene was placed downstream from the strong lambda phage promoter, PR, SEB was synthesized at readily detectable levels in E. coli. Interestingly, mature SEB was almost exclusively present in the cytoplasmic fraction. The SEB precursor was found associated with the cell membrane. The entB gene was introduced back into S. aureus, and the clones were shown to produce SEB. The entB gene has been located to a 2.1-kilobase-pair region. Maxam-Gilbert sequencing of a part of the entB gene yielded a DNA sequence that corresponds to the known amino acid sequence of SEB. Southern hybridization experiments showed that the entB gene was present on identical restriction fragments in the chromosomes of SEB-producer strains. The entB gene is absent from SEB-nonproducer strains.