Perkins J B, Bower S, Howitt C L, Yocum R R, Pero J
OmniGene Bioproducts, Inc., Cambridge, Massachusetts 02138, USA.
J Bacteriol. 1996 Nov;178(21):6361-5. doi: 10.1128/jb.178.21.6361-6365.1996.
Northern (RNA) blot analysis of the Bacillus subtilis biotin operon, bioWAFDBIorf2, detected at least two steady-state polycistronic transcripts initiated from a putative vegetative (Pbio) promoter that precedes the operon, i.e., a full-length 7.2-kb transcript covering the entire operon and a more abundant 5.1-kb transcript covering just the first five genes of the operon. Biotin and the B. subtilis birA gene product regulated synthesis of the transcripts. Moreover, replacing the putative Pbio promoter and regulatory sequence with a constitutive SP01 phage promoter resulted in higher-level constitutive synthesis. Removal of a rho-independent terminator-like sequence located between the fifth (bioB) and sixth (bioI) genes prevented accumulation of the 5.1-kb transcript, suggesting that the putative terminator functions to limit expression of bioI, which is thought to be involved in an early step in biotin synthesis.
对枯草芽孢杆菌生物素操纵子bioWAFDBIorf2进行的Northern(RNA)印迹分析检测到,至少有两种稳态多顺反子转录本是从操纵子之前一个假定的营养型(Pbio)启动子起始的,即一种全长7.2 kb的转录本覆盖整个操纵子,以及一种更为丰富的5.1 kb转录本仅覆盖操纵子的前五个基因。生物素和枯草芽孢杆菌birA基因产物调节这些转录本的合成。此外,用组成型SP01噬菌体启动子取代假定的Pbio启动子和调控序列会导致更高水平组成型合成。去除位于第五个(bioB)和第六个(bioI)基因之间的一个类rho非依赖性终止子序列会阻止5.1 kb转录本的积累,这表明假定的终止子起到限制bioI表达的作用,据认为bioI参与生物素合成的早期步骤。