温真;杨彩云;林丽玉;毛惠民;杨梅;

• 1:福建师范大学生命科学学院

摘要(Abstract)：

根据已报道的aiiA基因序列(AY460124)设计特异性PCR引物,扩增得到aiiA基因启动子序列的单一产物.将该启动子序列代替pET-28a-GFP质粒上的T7启动子,构建绿色荧光蛋白功能分析载体.经测序比对和生物信息学软件分析发现该序列含有6个可能的启动区域和19个不同类型的转录结合位点.最后通过荧光显微镜观察到绿色荧光蛋白发出荧光,从而分析了该序列的启动子活性.

关键词(KeyWords)： 苏云金芽胞杆菌;启动子;绿色荧光蛋白;功能鉴定

Abstract:

Keywords:

基金项目(Foundation): 福建省科技厅资助项目(2009N0032);; 福建省教育厅资助项目(JA08041)

作者(Authors): 温真;杨彩云;林丽玉;毛惠民;杨梅;

参考文献(References)：

• [1]Yeon K M,Cheong W S,Oh H S,et al.Quorum sensing:a new biofouling control paradigm in a membranebioreactor for advanced wastewater treatment[J].Environ Sci Technol,2009,43(2):380-385.
• [2]陈静,刘显军,边连全,等.细菌生物膜及细菌群体感应系统的研究进展[J].江苏农业科学,2010(5):32-33.
• [3]Von Bodman S B,Bauer W D,Coplin D L.Quorum sensing in plant-pathogenic bacteria[J].Annu RevPhytopathol,2003,41:455-482.
• [4]Pompeani A J,Irgon J J,Berger M F,et al.The Vibrio harveyi master quorum-sensing regulator,LuxR,aTetR-type protein is both an activator and a repressor:DNA recognition and binding specificity at target promoters[J].Mol Microbiol,2008,70(1):76-88.
• [5]Nackerdien Z E,Keynan A,Bassler B L,et al.Quorum sensing influences Vibrio harveyi growth rates in amanner not fully accounted for by the marker effect of bioluminescence[J].PLoS One,2008,3(2):e1671.
• [6]Vikstrom E,Tafazoli F,Magnusson K E.Pseudomonas aeruginosa quorum sensing molecule N-(3oxodod-ecan-oyl)-l-homoserine lactone disrupts epithelial barrier integrity of Caco-2 cells[J].FEBS Lett,2006,580(30):6921-6928.
• [7]Abbas A,Adams C,Scully N,et al.A role for TonB1in biofilm formation and quorum sensing in Pseudomonasaeruginosa[J].FEMS Microbiol Lett,2007,274(2):269-278.
• [8]Dong Y H,Gusti A R,Zhang Q,et al.Identification of quorum-quenching N-acyl homoserine lactonases fromBacillus species[J].Appl Environ Microbiol,2002,68(4):1754-1759.
• [9]Lee S J,Park S Y,Lee J J,et al.Genes encoding the N-acyl homoserine lactone-degrading enzyme are widespreadin many subspecies of Bacillus thuringiensis[J].Appl Environ Microbiol,2002,68(8):3919-3924.
• [10]Henke J M,Bassler B L.Three parallel quorum-sensing systems regulate gene expression in Vibrio harveyi[J].J Bacteriol,2004,186(20):6902-6914.
• [11]杨梅,姚帆,黄勤清,等.不同来源的苏云金芽孢杆菌aiiA基因表达及其抗病性分析[J].应用与环境生物学报,2007,13(3):373-381.
• [12]Dong Y H,Xu J L,Li X Z,et al.AiiA,an enzyme that inactivates the acylhomoserine lactone quorum-sensingsignal and attenuates the virulence of Erwinia carotovora[J].Proc Natl Acad Sci USA,2000,97(7):3526-3531.
• [13]Dong Y H,Wang L H,Xu J L,et al.Quenching quorum-sensing-dependent bacterial infection by an N-acylhomoserine lactonase[J].Nature,2001,411(6839):813-817.
• [14]Ulrich R L.Quorum quenching:enzymatic disruption of N-acylhomoserine lactone-mediated bacterial communic-ation in Burkholderia thailandensis[J].Appl Environ Microbiol,2004,70(10):6173-6180.
• [15]Molina L,Constantinescu F,Michel L,et al.Degradation of pathogen quorum-sensing molecules by soilbacteria:a preventive and curative biological control mechanism[J].FEMS Microbiol Ecol,2003,45(1):71-81.
• [16]Reimmann C,Ginet N,Michel L,et al.Genetically programmed autoinducer destruction reduces virulence geneexpression and swarming motility in Pseudomonas aeruginosa PAO1[J].Microbiology,2002,148:923-932.
• [17]Zhang L,Ruan L,Hu C,et al.Fusion of the genes for AHL-lactonase and S-layer protein in Bacillus thuring-iensis increases its ability to inhibit soft rot caused by Erwinia carotovora[J].Appl Microbiol Biotechnol,2007,74(3):667-675.
• [18]郝福英,周先碗.生物化学与分子生物学实验[M].北京:高等教育出版社,2009:7.