Predich M, Nair G, Smith I
Department of Microbiology, Public Health Research Institute, New York, New York.
J Bacteriol. 1992 May;174(9):2771-8. doi: 10.1128/jb.174.9.2771-2778.1992.
The Bacillus subtilis genes kinA (spoIIJ), spo0F, and spo0A encode components of the sporulation signal transduction pathway. Recent work has suggested that these genes are transcribed by a minor form of RNA polymerase, E sigma H (sigma H is the product of spo0H, another early sporulation gene). We directly tested this hypothesis by performing in vitro transcription assays with reconstituted E sigma H and a set of plasmids containing the kinA, spo0F, and spo0A promoter regions. We were able to obtain distinct transcripts of the expected sizes with all three genes by using linearized or supercoiled templates. Furthermore, primer extension experiments indicate that the transcription start sites for the three genes in vitro and in vivo are the same. In addition, we measured steady-state levels of kinA, spo0F, and spo0A mRNAs during growth in sporulation medium; all of them were increased at or near the beginning of the stationary phase.
枯草芽孢杆菌基因kinA（spoIIJ）、spo0F和spo0A编码芽孢形成信号转导途径的组分。最近的研究表明，这些基因由一种次要形式的RNA聚合酶E sigma H转录（sigma H是另一个早期芽孢形成基因spo0H的产物）。我们通过用重组的E sigma H和一组含有kinA、spo0F和spo0A启动子区域的质粒进行体外转录试验，直接验证了这一假设。通过使用线性化或超螺旋模板，我们能够获得这三个基因预期大小的不同转录本。此外，引物延伸实验表明，这三个基因在体外和体内的转录起始位点是相同的。另外，我们测量了在芽孢形成培养基中生长期间kinA、spo0F和spo0A mRNA的稳态水平；它们在稳定期开始时或接近稳定期开始时均增加。
