Brakhage A A, Browne P, Turner G
Department of Molecular Biology and Biotechnology, University of Sheffield, United Kingdom.
J Bacteriol. 1992 Jun;174(11):3789-99. doi: 10.1128/jb.174.11.3789-3799.1992.
Expression of the Aspergillus nidulans penicillin biosynthesis genes acvA and ipnA, encoding delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase and isopenicillin N synthetase, respectively, was analyzed. The intergenic region carrying the divergently oriented promoters was fused in frame in both orientations to Escherichia coli lacZ and E. coli uidA reporter genes. Each construct permits simultaneous expression studies of both genes. Transformants of A. nidulans carrying a single copy of either plasmid integrated at the chromosomal argB locus were selected for further investigations. Expression of both genes was directed by the 872-bp intergenic region. ipnA- and acvA-derived gene fusions were expressed from this region at different levels. ipnA had significantly higher expression than did acvA. Glucose specifically reduced the production of penicillin and significantly repressed the expression of ipnA but not of acvA gene fusions. The specific activities of isopenicillin N synthetase, the gene product of ipnA, and acyl coenzyme A:6-aminopenicillanic acid acyltransferase were also reduced in glucose-grown cultures.
对构巢曲霉青霉素生物合成基因acvA和ipnA的表达进行了分析,这两个基因分别编码δ-(L-α-氨基己二酰基)-L-半胱氨酰-D-缬氨酸合成酶和异青霉素N合成酶。携带反向排列启动子的基因间区域在两个方向上均与大肠杆菌lacZ和大肠杆菌uidA报告基因进行了读框融合。每个构建体都允许对这两个基因进行同步表达研究。选择在染色体argB位点整合了单个质粒拷贝的构巢曲霉转化子进行进一步研究。两个基因的表达均由872 bp的基因间区域指导。源自ipnA和acvA的基因融合体在该区域以不同水平表达。ipnA的表达明显高于acvA。葡萄糖特异性降低了青霉素的产量,并显著抑制了ipnA基因融合体的表达,但对acvA基因融合体没有影响。在葡萄糖培养的菌体中,ipnA的基因产物异青霉素N合成酶以及酰基辅酶A:6-氨基青霉烷酸酰基转移酶的比活性也降低了。