Feng B, Friedlin E, Marzluf G A
Department of Biochemistry, Ohio State University, Columbus 43210.
Appl Environ Microbiol. 1994 Dec;60(12):4432-9. doi: 10.1128/aem.60.12.4432-4439.1994.
Vectors which possess a truncated niaD gene encoding nitrate reductase were developed to allow targeted gene integration during transformation of an niaD mutant Penicillium chrysogenum host. The Penicillium genes pcbC and penAB are immediately adjacent to each other and are divergently transcribed, with an intergenic control region serving as their promoters. Gene fusions were constructed with a reporter gene, uidA, which encodes beta-glucuronidase. The pcbC-penAB intergenic region was fused to the uidA gene in both orientations so that regulated expression of each structural gene could be investigated. These fusion genes were targeted to the chromosomal site of the niaD locus of P. chrysogenum, and their expression was examined under different growth conditions. The expression of each of these penicillin biosynthesis genes was found to be regulated by nitrogen repression, glucose repression, and growth stage control.
构建了携带编码硝酸还原酶的截短niaD基因的载体,以便在niaD突变型产黄青霉宿主转化过程中实现靶向基因整合。产黄青霉基因pcbC和penAB彼此紧邻且转录方向相反,一个基因间控制区域作为它们的启动子。用编码β-葡萄糖醛酸酶的报告基因uidA构建了基因融合体。将pcbC-penAB基因间区域以两种方向与uidA基因融合,从而能够研究每个结构基因的调控表达。这些融合基因被靶向到产黄青霉niaD基因座的染色体位点,并在不同生长条件下检测它们的表达。发现这些青霉素生物合成基因中的每一个的表达都受氮阻遏、葡萄糖阻遏和生长阶段控制。