通过改进枯草芽孢杆菌中青色荧光蛋白和黄色荧光蛋白的产生来可视化差异基因表达
Visualization of differential gene expression by improved cyan fluorescent protein and yellow fluorescent protein production in Bacillus subtilis.
作者信息
Veening Jan-Willem, Smits Wiep Klaas, Hamoen Leendert W, Jongbloed Jan D H, Kuipers Oscar P
机构信息
Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Haren, The Netherlands.
出版信息
Appl Environ Microbiol. 2004 Nov;70(11):6809-15. doi: 10.1128/AEM.70.11.6809-6815.2004.
Abstract
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.
摘要
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。
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本文引用的文献
1
The Spo0A regulon of Bacillus subtilis.枯草芽孢杆菌的Spo0A调控子。
Mol Microbiol. 2003 Dec;50(5):1683-701. doi: 10.1046/j.1365-2958.2003.03818.x.
2
Cannibalism by sporulating bacteria.产孢细菌的自相残杀现象。
Science. 2003 Jul 25;301(5632):510-3. doi: 10.1126/science.1086462. Epub 2003 Jun 19.
3
The Bacillus subtilis transition state regulator AbrB binds to the -35 promoter region of comK.枯草芽孢杆菌过渡态调节因子AbrB与comK的 -35启动子区域结合。
FEMS Microbiol Lett. 2003 Jan 28;218(2):299-304. doi: 10.1111/j.1574-6968.2003.tb11532.x.
4
Stochastic gene expression in a single cell.单细胞中的随机基因表达。
Science. 2002 Aug 16;297(5584):1183-6. doi: 10.1126/science.1070919.
5
Cell cycle-dependent localization of two novel prokaryotic chromosome segregation and condensation proteins in Bacillus subtilis that interact with SMC protein.两种与SMC蛋白相互作用的新型原核生物染色体分离和凝聚蛋白在枯草芽孢杆菌中的细胞周期依赖性定位。
EMBO J. 2002 Jun 17;21(12):3108-18. doi: 10.1093/emboj/cdf314.
6
Construction and application of epitope- and green fluorescent protein-tagging integration vectors for Bacillus subtilis.枯草芽孢杆菌表位和绿色荧光蛋白标签整合载体的构建与应用
Appl Environ Microbiol. 2002 May;68(5):2624-8. doi: 10.1128/AEM.68.5.2624-2628.2002.
7
Regulation of noise in the expression of a single gene.单个基因表达中的噪声调控。
Nat Genet. 2002 May;31(1):69-73. doi: 10.1038/ng869. Epub 2002 Apr 22.
8
A ComGA-dependent checkpoint limits growth during the escape from competence.一种依赖ComGA的检查点在从感受态逃逸过程中限制生长。
Mol Microbiol. 2001 Apr;40(1):52-64. doi: 10.1046/j.1365-2958.2001.02363.x.
9
Improved plasmid vectors for the production of multiple fluorescent protein fusions in Bacillus subtilis.用于在枯草芽孢杆菌中生产多种荧光蛋白融合体的改良质粒载体。
Gene. 2001 Feb 21;264(2):289-97. doi: 10.1016/s0378-1119(01)00338-9.
10
Control of sporulation initiation in Bacillus subtilis.枯草芽孢杆菌中孢子形成起始的调控
Curr Opin Microbiol. 2000 Dec;3(6):561-6. doi: 10.1016/s1369-5274(00)00141-7.
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