Jackson R, Cram D, Ray A, DiBerardino D, Skurray R
Mol Gen Genet. 1984;197(1):129-36. doi: 10.1007/BF00327933.
We describe the molecular cloning of BglII fragments of the hybrid plasmid pRS5 (pSC101 and EcoRI fragments of F; f7, f5, f3 and f6). The clones isolated were examined for the expression of F-specified replication, incompatibility, mobilization and inhibition of T7 bacteriophage multiplication. Proteins directed by the BglII clones were labelled in Escherichia coli K12 maxicells and analyzed by SDS-polyacrylamide gel electrophoresis. The sizes of previously reported proteins, encoded by the replication, incompatibility and leading regions encompassed by these plasmids have been confirmed in this study. In addition, the results demonstrate that a pif gene, which encodes an 80,000 dalton polypeptide essential for the inhibition T7 phage multiplication, is located on the BglII fragment that spans the junction of EcoRI fragments f7 and f5.
我们描述了杂交质粒pRS5（pSC101和F的EcoRI片段；f7、f5、f3和f6）的BglII片段的分子克隆。对分离得到的克隆进行了F特异性复制、不相容性、迁移以及对T7噬菌体增殖抑制的表达检测。由BglII克隆指导合成的蛋白质在大肠杆菌K12大细胞中进行标记，并通过SDS-聚丙烯酰胺凝胶电泳进行分析。本研究证实了这些质粒所包含的复制、不相容性和先导区域编码的先前报道的蛋白质的大小。此外，结果表明，一个编码对抑制T7噬菌体增殖至关重要的80000道尔顿多肽的pif基因，位于跨越EcoRI片段f7和f5连接处的BglII片段上。
