Onyeocha I, Vieille C, Zimmer W, Baca B E, Flores M, Palacios R, Elmerich C
URA 1300 CNRS, Département des Biotechnologies, Institut Pasteur, Paris, France.
Plasmid. 1990 May;23(3):169-82. doi: 10.1016/0147-619x(90)90049-i.
Homology was previously detected between the DNA restriction fragments containing Rhizobium meliloti nodulation genes and the 90-MDa plasmid, p90, of Azospirillum brasilense Sp7. Two DNA loci from Sp7 genome that complement mutations in the exopolysaccharide synthesis genes, exoB and exoC, of R. meliloti were also shown to be present on the plasmid. A more detailed characterization of the plasmid was undertaken to establish its physical map and to localize the nod homologies and other specific regions. Six loci were mapped, the region homologous to the nodulation genes, nodPQ, of R. meliloti, the exoB and exoC mutation-correcting loci, a locus for Ap resistance, a bla homology region different from the Ap resistance locus, and a region necessary for the maintenance of p90 as an independent replicon. Mobilization into Agrobacterium tumefaciens of p90-Tn5-Mob was obtained at a frequency of 10(-4), with the plasmid helper pJB3JI. Self-transfer of p90 was not demonstrated. Fragments of p90 hybridized with a plasmid of 90 MDa present in most A. brasilense and some A. lipoferum strains, suggesting a plasmid family in Azospirillum.
先前已检测到,含有苜蓿根瘤菌结瘤基因的DNA限制性片段与巴西固氮螺菌Sp7的90-MDa质粒p90之间存在同源性。还发现,来自Sp7基因组的两个DNA位点可互补苜蓿根瘤菌胞外多糖合成基因exoB和exoC中的突变,这两个位点也存在于该质粒上。为了构建其物理图谱并定位结瘤同源性区域和其他特定区域,对该质粒进行了更详细的表征。绘制了六个位点的图谱,包括与苜蓿根瘤菌结瘤基因nodPQ同源的区域、exoB和exoC突变校正位点、一个抗氨苄青霉素位点、一个与抗氨苄青霉素位点不同的bla同源区域,以及p90作为独立复制子维持所必需的区域。利用质粒辅助载体pJB3JI,p90-Tn5-Mob以10(-4)的频率转入根癌土壤杆菌。未证明p90能自我转移。p90的片段与大多数巴西固氮螺菌和一些产脂固氮螺菌菌株中存在的90 MDa质粒杂交,表明在固氮螺菌中存在一个质粒家族。