Imanaka T, Himeno T, Aiba S
J Bacteriol. 1987 Sep;169(9):3867-72. doi: 10.1128/jb.169.9.3867-3872.1987.
The penicillinase antirepressor gene, penJ, of Bacillus licheniformis ATCC 9945a was cloned in Escherichia coli by using pMB9 as a vector plasmid. The penicillinase gene, penP, its repressor gene, penI, and penJ were encoded on the cloned 5.2-kilobase HindIII fragment of the recombinant plasmid pTTE71. The penJ open reading frame was composed of 1,803 bases and 601 amino acid residues (molecular weight, 68,388). A Shine-Dalgarno sequence was found 7 bases upstream from the translation start site. Since this sequence was located in the 3'-terminal region of the penI gene, penJ might be transcribed together with penI as a polycistronic mRNA from the penI promoter. Frameshift mutations of penJ were constructed in vitro from pTTE71, and the penJ mutant gene was introduced into B. licheniformis by chromosomal recombination. The transformant B. licheniformis U173 (penP+ penI+ penJ) turned out to be uninducible for penicillinase production, whereas the wild-type strain (penP+ penI+ penJ+) was inducible. Only when these three genes (penP, penI, and PenJ) were simultaneously subcloned in Bacillus subtilis did the plasmid carrier exhibit inducible penicillinase production, as did wild-type B. licheniformis. It was concluded that penJ is involved in the penicillinase induction. The regulation of penP expression by penI and penJ is discussed.
地衣芽孢杆菌ATCC 9945a的青霉素酶抗阻遏基因penJ，以pMB9作为载体质粒在大肠杆菌中进行了克隆。青霉素酶基因penP、其阻遏基因penI和penJ编码于重组质粒pTTE71克隆的5.2千碱基HindIII片段上。penJ开放阅读框由1803个碱基和601个氨基酸残基组成（分子量为68388）。在翻译起始位点上游7个碱基处发现了一个Shine-Dalgarno序列。由于该序列位于penI基因的3'末端区域，penJ可能与penI一起从penI启动子转录为多顺反子mRNA。从pTTE71体外构建了penJ的移码突变体，并通过染色体重组将penJ突变基因导入地衣芽孢杆菌。转化后的地衣芽孢杆菌U173（penP+ penI+ penJ）对青霉素酶的产生无诱导性，而野生型菌株（penP+ penI+ penJ+）具有诱导性。只有当这三个基因（penP、penI和PenJ）同时亚克隆到枯草芽孢杆菌中时，质粒载体才表现出与野生型地衣芽孢杆菌一样的诱导性青霉素酶产生。得出的结论是，penJ参与青霉素酶的诱导。讨论了penI和penJ对penP表达的调控。
