Díez B, Gutiérrez S, Barredo J L, van Solingen P, van der Voort L H, Martín J F
Department of Ecology, Genetics and Microbiology, University of León, Spain.
J Biol Chem. 1990 Sep 25;265(27):16358-65.
Penicillium chrysogenum DNA fragments cloned in EMBL3 or cosmid vectors from the upstream region of the pcbC-penDE cluster carry a gene (pcbAB) that complemented the deficiency of alpha-aminoadipyl-cysteinyl-valine synthetase of mutants npe5 and npe10, and restored penicillin production to mutant npe5. A protein of about 250 kDa was observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels of cell-free extracts of complemented strains that was absent in the npe5 and npe10 mutants but exists in the parental strain from which the mutants were obtained. Transcriptional mapping studies showed the presence of one long transcript of about 11.5 kilobases that hybridized with several probes internal to the pcbAB gene, and two small transcripts of 1.15 kilobases that hybridized with the pcbC or the penDE gene, respectively. The transcription initiation and termination regions of the pcbAB gene were mapped by hybridization with several small probes. The region has been completely sequenced. It includes an open reading frame of 11,376 nucleotides that encodes a protein with a deduced Mr of 425,971. Three repeated dominia were found in the alpha-aminoadipyl-cysteinyl-valine synthetase which have high homology with the gramicidin synthetase I and tyrocidine synthetase I. The pcbAB is linked to the pcbC and penDE genes and is transcribed in the opposite orientation to them.
从产黄青霉（Penicillium chrysogenum）的pcbC - penDE基因簇上游区域克隆到EMBL3或黏粒载体中的DNA片段携带一个基因（pcbAB），该基因可弥补npe5和npe10突变体α-氨基己二酰-半胱氨酰-缬氨酸合成酶的缺陷，并使npe5突变体恢复青霉素生产。在互补菌株的无细胞提取物的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳凝胶中观察到一种约250 kDa的蛋白质，该蛋白质在npe5和npe10突变体中不存在，但存在于获得这些突变体的亲本菌株中。转录图谱研究表明存在一个约11.5千碱基的长转录本，它与pcbAB基因内部的几个探针杂交，还有两个分别为1.15千碱基的小转录本，它们分别与pcbC或penDE基因杂交。通过与几个小探针杂交绘制了pcbAB基因的转录起始和终止区域。该区域已被完全测序。它包括一个11376个核苷酸的开放阅读框，编码一种推导的Mr为425971的蛋白质。在α-氨基己二酰-半胱氨酰-缬氨酸合成酶中发现了三个重复结构域，它们与短杆菌肽合成酶I和短杆菌酪肽合成酶I具有高度同源性。pcbAB与pcbC和penDE基因相连，并以与它们相反的方向转录。
