Rossbach S, Schell J, de Bruijn F J
Max-Planck-Institut für Züchtungsforschung, Abteilung Genetische Grundlagen der Pflanzenzüchtung, D-5000, Köln 30, Federal Republic of Germany.
Mol Gen Genet. 1987 Oct;209(3):419-26. doi: 10.1007/BF00331144.
The ntrC locus of Agrobacterium tumefaciens C58 has been cloned using the Azorhizobium sesbaniae ORS571 ntrC gene as a DNA hybridization probe. Transposon Tn5 mutagenesis of the cloned ntrC locus was carried out and one Tn5 insertion within the region of highest DNA homology with A. sesbaniae ORS571 ntrC was used for gene replacement of the wild-type C58 ntrC gene. The A. tumefaciens ntrC::Tn5 mutant was found to be unable to grow on nitrate as sole nitrogen (N) source, to lack glutamine synthetase (GSII) activity and to be unable to use arginine (or ornithine) as sole N source, unless the Ti-encoded arginine catabolism pathway was induced with small amounts of nopaline. Thus the A. tumefaciens ntrC regulatory gene is essential for (transcriptional) activation of the GSII and nitrate reductase genes, as well as for the chromosomal but not the Ti-borne arginine catabolism pathways.
利用Sesbania根瘤菌ORS571的ntrC基因作为DNA杂交探针,克隆了根癌农杆菌C58的ntrC基因座。对克隆的ntrC基因座进行转座子Tn5诱变,并将与Sesbania根瘤菌ORS571 ntrC具有最高DNA同源性区域内的一个Tn5插入用于野生型C58 ntrC基因的基因置换。发现根癌农杆菌ntrC::Tn5突变体不能以硝酸盐作为唯一氮源生长,缺乏谷氨酰胺合成酶(GSII)活性,并且不能以精氨酸(或鸟氨酸)作为唯一氮源,除非用少量胭脂碱诱导Ti编码的精氨酸分解代谢途径。因此,根癌农杆菌ntrC调控基因对于GSII和硝酸还原酶基因的(转录)激活以及染色体上而非Ti携带的精氨酸分解代谢途径至关重要。