Komai N, Nishizawa T, Hayakawa Y, Murotsu T, Matsubara K
Mol Gen Genet. 1982;186(2):193-203. doi: 10.1007/BF00331850.
Various DNA subfragments were derived from miniF DNA by complete or partial PstI cleavage, and cloned in the plasmid vectors pBR322 or lambda dv1. The recombinant plasmids obtained were introduced into an Escherichia coli minicell-producing strain, and the plasmid-coded proteins were radiolabeled and analyzed by gel electrophoresis. Six miniF-encoded proteins, larger than 11 000 daltons, were detected and their coding regions were mapped on the F plasmid genome. Three of them were assigned by taking into account the known nucleotide sequences (Murotsu et al. 1981; K. Yoshioka, personal communication). The coding directions of some proteins were determined by inserting the lac promotor into one of the recombinant plasmids and analyzing the increase in production of the proteins. The coding direction of the five proteins analyzed so far was uniform. Comparison of these results with a functional map of miniF suggested possible roles of the proteins.
通过完全或部分PstI切割从miniF DNA获得各种DNA亚片段，并克隆到质粒载体pBR322或λdv1中。将获得的重组质粒导入产大肠杆菌小细胞的菌株中，对质粒编码的蛋白质进行放射性标记并通过凝胶电泳分析。检测到六种大于11000道尔顿的miniF编码蛋白，并将它们的编码区域定位在F质粒基因组上。其中三种是通过考虑已知的核苷酸序列确定的（Murotsu等人，1981年；K. Yoshioka，个人交流）。通过将lac启动子插入其中一个重组质粒并分析蛋白质产量的增加来确定一些蛋白质的编码方向。到目前为止分析的五种蛋白质的编码方向是一致的。将这些结果与miniF的功能图谱进行比较，提示了这些蛋白质可能的作用。
