Veening Jan-Willem, Smits Wiep Klaas, Hamoen Leendert W, Jongbloed Jan D H, Kuipers Oscar P
Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Haren, The Netherlands.
Appl Environ Microbiol. 2004 Nov;70(11):6809-15. doi: 10.1128/AEM.70.11.6809-6815.2004.
The distinguishable cyan and yellow fluorescent proteins (CFP and YFP) enable the simultaneous in vivo visualization of different promoter activities. Here, we report new cloning vectors for the construction of cfp and yfp fusions in Bacillus subtilis. By extending the N-terminal portions of previously described CFP and YFP variants, 20- to 70-fold-improved fluorescent-protein production was achieved. Probably, the addition of sequences encoding the first eight amino acids of the N-terminal part of ComGA of B. subtilis overcomes the slow translation initiation that is provoked by the eukaryotic codon bias present in the original cfp and yfp genes. Using these new vectors, we demonstrate that, within an isogenic population of sporulating B. subtilis cells, expression of the abrB and spoIIA genes is distinct in individual cells.
可区分的青色和黄色荧光蛋白（CFP和YFP）能够在体内同时可视化不同的启动子活性。在此，我们报告了用于在枯草芽孢杆菌中构建CFP和YFP融合体的新型克隆载体。通过延伸先前描述的CFP和YFP变体的N端部分，荧光蛋白产量提高了20至70倍。可能是，添加编码枯草芽孢杆菌ComGA N端部分前八个氨基酸的序列克服了原始cfp和yfp基因中存在的真核密码子偏好所引发的缓慢翻译起始。使用这些新载体，我们证明，在产孢枯草芽孢杆菌细胞的同基因群体中，abrB和spoIIA基因在单个细胞中的表达是不同的。
