Smith A A, Greene R C, Kirby T W, Hindenach B R
Proc Natl Acad Sci U S A. 1985 Sep;82(18):6104-8. doi: 10.1073/pnas.82.18.6104.
We have modified a previously isolated metJ plasmid by removing a segment of DNA including the rop gene. Bacterial strains carrying this plasmid produce elevated levels of the metJ gene product, presumably because of the high number of gene copies in the cell. We have isolated the metJ gene product in nearly homogeneous form from such a strain. The subunit size and the amino acid composition are the same as those predicted from the DNA sequence of the metJ gene. Sedimentation equilibrium measurements show that the native metJ gene product is a dimer. The purified dimer protects a short segment of DNA in the regulatory region of the metB and metJ genes from hydrolysis by DNase I.
我们通过去除一段包含rop基因的DNA片段,对先前分离出的metJ质粒进行了改造。携带该质粒的细菌菌株产生了高水平的metJ基因产物,推测这是由于细胞中基因拷贝数众多。我们已从这样的菌株中以近乎纯一的形式分离出了metJ基因产物。其亚基大小和氨基酸组成与根据metJ基因的DNA序列预测的结果相同。沉降平衡测量表明,天然的metJ基因产物是二聚体。纯化后的二聚体可保护metB和metJ基因调控区域中的一小段DNA不被DNase I水解。