Katzy E I, Matora L Y, Serebrennikova O B, Scheludko A V
Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, prospekt Entuziastov 13, Saratov, 410015, Russia.
Plasmid. 1998 Jul;40(1):73-83. doi: 10.1006/plas.1998.1353.
Using Omegon-Km mutagenesis, six Azospirillum brasilense Sp245 mutant derivatives lacking the capability to synthesize either one of the two major O-specific polysaccharides (O-PSs) were constructed in vivo. In all of the Lps mutants obtained, single Omegon-Km insertions were shown to be located on an indigenous plasmid DNA with molecular weight 120 MDa (p120). Physical and immunochemical analyses revealed two p120 loci coding for O-PSI and two p120 loci involved in the production of O-PSII. One of the lps loci from both groups was also shown to act in the production of Calcofluor-binding polysaccharides. It was demonstrated that two Sp245 plasmid bands with apparent molecular weights of 120 and 130 MDa (as visualized by analytical gel electrophoreses) seem to be the two topological forms of the same plasmid species (p120). Transfer properties of p120 were also examined.
利用奥米贡 - Km诱变技术,在体内构建了六株巴西固氮螺菌Sp245突变衍生物,它们缺乏合成两种主要O - 特异性多糖(O - PSs)中任何一种的能力。在所有获得的Lps突变体中,单个奥米贡 - Km插入片段显示位于分子量为120 MDa的内源质粒DNA(p120)上。物理和免疫化学分析揭示了两个编码O - PSI的p120位点以及两个参与O - PSII产生的p120位点。两组中的一个lps位点也被证明在钙黄绿素结合多糖的产生中起作用。结果表明,两条表观分子量分别为120和130 MDa的Sp245质粒条带(通过分析凝胶电泳观察)似乎是同一质粒种类(p120)的两种拓扑形式。还研究了p120的转移特性。