Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
Plasmid. 2011 Jul;66(2):85-92. doi: 10.1016/j.plasmid.2011.06.001. Epub 2011 Jun 25.
RNA transcripts from 199-kb incompatibility P-7 plasmid pCAR1 were analyzed using microarrays with evenly tiled probes with a nine-nucleotide offset in six different Pseudomonas host strains. We re-annotated 12 ORFs based on their RNA maps and on the comparisons with other sequenced IncP-7 plasmids. Ninety six of two hundred ORFs were identified on the IncP-7 backbone related to basic functions of the plasmid (replication, partition and conjugative transfer). More than 90% ORFs on the backbone were transcribed in each host strain, suggesting their importance for the plasmid survival in the host strains. Genes related to partition and conjugative transfer were differentially transcribed host by host, whereas the repA gene encoding replication initiation protein was transcribed at comparable level in each host. As for other plasmid 'accessory genes' of pCAR1 encoding carbazole degradation, putative transporter, or transposase were also differentially transcribed among different host strains. These differences may have resulted in distinct behaviors of the plasmid or of its host strain, and RNA maps of pCAR1 give us important information to understand the plasmid behaviors in different environments.
使用带有九个核苷酸偏移的均匀平铺探针的微阵列分析了来自 199-kb 不兼容 P-7 质粒 pCAR1 的 RNA 转录物,在六个不同的假单胞菌宿主菌株中进行了分析。我们根据 RNA 图谱和与其他已测序的 IncP-7 质粒的比较,重新注释了 12 个 ORF。在 IncP-7 骨架上鉴定出了 200 个 ORF 中的 96 个,这些 ORF 与质粒的基本功能(复制、分区和共轭转移)有关。在每个宿主菌株中,骨架上超过 90%的 ORF 都被转录,这表明它们对质粒在宿主菌株中的存活很重要。与分区和共轭转移相关的基因在宿主之间存在差异转录,而编码复制起始蛋白的 repA 基因在每个宿主中的转录水平相当。对于 pCAR1 的其他质粒“辅助基因”,编码咔唑降解、假定转运蛋白或转座酶的基因也在不同的宿主菌株中存在差异转录。这些差异可能导致质粒或其宿主菌株的行为不同,而 pCAR1 的 RNA 图谱为我们提供了理解质粒在不同环境中的行为的重要信息。