Fernández-López M, D'Haeze W, Mergaert P, Verplancke C, Promé J C, Van Montagu M, Holsters M
Laboratorium voor Genetica, Flanders Interuniversity Institute for Biotechnology, Universiteit Gent, Belgium.
Mol Microbiol. 1996 Jun;20(5):993-1000. doi: 10.1111/j.1365-2958.1996.tb02540.x.
Lipo-chitooligosaccharide (LCO) Nod factors are produced and secreted by rhizobia and trigger nodule development in leguminous host plants. The products of the bacterial nodlJ genes are related to transporters of capsular polysaccharides and were proposed to be involved in LCO transport. We have studied nodlJ of Azorhizobium caulinodans ORS571 by analysis of cell-associated and secreted radioactively labelled Nod factors in wild-type ORS571, a nodJ mutant and a complemented strain. Secretion was strongly reduced in the nodJ mutant, and restored to wild-type levels after complementation. Constructs were made for expression of combinations of different nod genes in Escherichia coli DH5 alpha. The strain DH5 alpha (pUCNABCSU) synthesized LCOs, but they were only secreted when a plasmid containing both nodl and nodJ was supplied in trans. nodl or nodJ alone was not sufficient. In E. coli as well as in Azorhizobium, the nodlJ-encoded transporter showed a specificity for more hydrophilic LCOs.
脂壳寡糖(LCO)结瘤因子由根瘤菌产生并分泌,可触发豆科宿主植物的根瘤发育。细菌nodlJ基因的产物与荚膜多糖转运蛋白有关,有人提出其参与LCO的转运。我们通过分析野生型ORS571、nodJ突变体和互补菌株中与细胞相关的和分泌的放射性标记结瘤因子,研究了茎瘤固氮根瘤菌ORS571的nodlJ。在nodJ突变体中分泌显著减少,互补后恢复到野生型水平。构建了用于在大肠杆菌DH5α中表达不同nod基因组合的载体。菌株DH5α(pUCNABCSU)合成了LCO,但只有在反式提供同时含有nodl和nodJ的质粒时才会分泌。单独的nodl或nodJ是不够的。在大肠杆菌和茎瘤固氮根瘤菌中,nodlJ编码的转运蛋白对亲水性更强的LCO具有特异性。