Replication of the Streptomyces plasmid pSN22 through single-stranded intermediates |
| |
Authors: | Masakazu Kataoka Norihito Kuno Takashi Horiguchi Tatsuji Seki Toshiomi Yoshida |
| |
Affiliation: | 1. International Center of Cooperative Research in Biotechnology, Japan 2. Faculty of Engineering, Osaka University, Yamada-oka 2-1, 565, Suita-shi, Osaka, Japan
|
| |
Abstract: | The replication of the 11 kb conjugative multicopy Streptomyces plasmid pSN22 was analyzed. Mutation and complementation analyses indicated that the minimal region essential for plasmid replication was located on a 1.9 kb fragment of pSN22, containing a trans-acting element encoding a replication protein and a cis-acting sequence acting as a replication origin. Southern hybridization showed that minimal replicon plasmids accumulated much more single-stranded plasmid molecules than did wild-type pSN22. Only one strand was accumulated. A 500 by fragment from the pSN22 transfer region was identified which reduced the relative amount of single-stranded DNA, when added in the native orientation to minimal replicon plasmids. This 500 by DNA sequence may be an origin for second-strand synthesis. It had no effect on the efficiency of co-transformation, plasmid incompatibility, or stability. The results indicate that pSN22 replicates via single-stranded intermediates by a rolling circle mechanism. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|