Geider K, Baldes R
Max-Planck-Institut für medizinische Forschung, Abteilung Molekulare Biologie, Heidelberg, FRG.
Nucleic Acids Res. 1988 Jul 25;16(14A):6385-96. doi: 10.1093/nar/16.14.6385.
Plasmids with the replication origin of bacteriophage fd, the pfd-plasmids, were investigated for compatibility in E. coli cells expressing fd gene 2-protein. This was measured by transformation of Ca-treated cells with and without a residing pfd-plasmid. When the two plasmids contained the complete intergenic region of bacteriophage fd, they were fully compatible in contrast to the situation in which at least one plasmid had a shortened origin for viral strand replication. This incompatibility effect was partially compensated for by a pfd-plasmid with a short origin and with the fd gene 2. The fd replication origin on a colEl plasmid did not affect compatibility in polA+ cells indicating its idling in the presence of the colEl origin. It can be concluded that a short replication origin requires high amounts of gene 2-protein in contrast to the long origin. Accumulation of replication intermediates severely interferes with host cell metabolism.
对具有噬菌体fd复制起点的质粒(即pfd质粒),在表达fd基因2蛋白的大肠杆菌细胞中进行了相容性研究。通过用含有或不含有常驻pfd质粒的Ca处理细胞进行转化来测定相容性。当两个质粒含有噬菌体fd完整的基因间区域时,它们完全相容,这与至少一个质粒具有缩短的病毒链复制起点的情况形成对比。这种不相容效应部分地被具有短起点和fd基因2的pfd质粒所补偿。在colE1质粒上的fd复制起点在polA+细胞中不影响相容性,表明其在存在colE1起点时处于闲置状态。可以得出结论,与长起点相比,短复制起点需要大量的基因2蛋白。复制中间体的积累严重干扰宿主细胞代谢。