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这对于线性质粒 pBSSB1 的稳定性是必不可少的,并且影响伤寒沙门氏菌血清型的运动性。

Fis is essential for the stability of linear plasmid pBSSB1 and affects the motility of Salmonella enterica serovar Typhi.

机构信息

Department of Biochemistry and Molecular Biology, School of Medical Technology, Jiangsu University, Zhenjiang, Jiangsu, China.

出版信息

PLoS One. 2012;7(7):e37462. doi: 10.1371/journal.pone.0037462. Epub 2012 Jul 23.

Abstract

pBSSB1 is a 27 kb non-bacteriophage-related linear plasmid first found in Salmonella enterica serovar Typhi (S. Typhi), but the mechanism underlying the replication of pBSSB1 is currently unknown. Previous reports showed that the factor for inversion stimulation (Fis) encoded by fis can affect the replication, transcription and other processes through binding DNA. Here, a fis deletion mutant of S. Typhi (Δfis) was prepared through the homologous recombination mediated by suicide plasmid and the loss of pBSSB1 in Δfis was observed surprisingly by pulsed field gel electrophoresis (PFGE). Subsequently, the loss of pBSSB1 was verified by PCR and Southern blot. In addition, the motility of Δfis was deficient and the flagellin of Δfis could not be detected by 2-dimensional polyacrylamide gel electrophoresis. All these results show that Fis is essential for the stability of pBSSB1 and affects the motility of S. Typhi.

摘要

pBSSB1 是一种 27kb 的非噬菌体相关线性质粒,最初在伤寒沙门氏菌(S. Typhi)中发现,但目前尚不清楚 pBSSB1 复制的机制。先前的报告表明,由 fis 编码的倒位刺激因子(Fis)可以通过结合 DNA 影响复制、转录和其他过程。在这里,通过自杀质粒介导的同源重组制备了伤寒沙门氏菌的 fis 缺失突变体(Δ fis),令人惊讶的是,脉冲场凝胶电泳(PFGE)观察到了 Δ fis 中 pBSSB1 的缺失。随后,通过 PCR 和 Southern blot 验证了 pBSSB1 的缺失。此外,Δ fis 的运动性缺陷,并且通过 2 维聚丙烯酰胺凝胶电泳无法检测到 Δ fis 的鞭毛蛋白。所有这些结果表明,Fis 对于 pBSSB1 的稳定性是必需的,并且影响伤寒沙门氏菌的运动性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12bb/3402438/980a7035730f/pone.0037462.g001.jpg

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