Gömpel-Klein P, Mack M, Brendel M
Institut für Mikrobiologie, Johann Wolfgang Goethe-Universität, Frankfurt/Main, Federal Republic of Germany.
Curr Genet. 1989 Aug;16(2):65-74. doi: 10.1007/BF00393397.
The genes SNQ and SFA confer hyperresistance to 4-NQO and FA when present on a multi-copy plasmid in yeast. Both are non-essential genes since transplacement of SNQ by a disrupted snq-0::LEU2 yielded stable and viable haploid integrants. Southern analysis revealed that SNQ and SFA are single-loci genes, and OFAGE analysis showed that they are located on chromosome XIII and IV, respectively. Northern blot analysis of SNQ and SFA revealed poly(A)+ RNA transcripts of 2 kb and 1.7 kb, respectively. Nuclease S1 mapping showed SNQ to have a coding region of 1.6 kb and SFA, one of 1.3 kb. The 5' coding regions were determined for both genes, while the 3' end could only be determined for gene SNQ. Both genes do not appear to contain introns. The SFA locus was also mapped by transposon mutagenesis. Tn10-LUK integrants disrupted the SFA gene function at sites that were determined by subcloning to lie within the SFA transcription unit.
当基因SNQ和SFA存在于酵母中的多拷贝质粒上时,可赋予对4-硝基喹啉氧化物(4-NQO)和氟乙酸(FA)的超抗性。由于用破坏的snq-0::LEU2对SNQ进行转座置换产生了稳定且有活力的单倍体整合体,所以这两个基因都是非必需基因。Southern分析表明SNQ和SFA是单基因座基因,而脉冲场交变凝胶电泳(OFAGE)分析表明它们分别位于第十三号和四号染色体上。对SNQ和SFA的Northern印迹分析分别揭示了2 kb和1.7 kb的聚腺苷酸化(poly(A)+)RNA转录本。核酸酶S1图谱显示SNQ有一个1.6 kb的编码区,SFA有一个1.3 kb的编码区。确定了两个基因的5'编码区,而只能确定基因SNQ的3'末端。两个基因似乎都不包含内含子。SFA基因座也通过转座子诱变进行了定位。Tn10-LUK整合体在通过亚克隆确定位于SFA转录单位内的位点破坏了SFA基因功能。