REGULATION OF THE ALGINATE BIOSYNTHESIS GENE ALGC IN PSEUDOMONAS-AERUGINOSA DURING BIOFILM DEVELOPMENT IN CONTINUOUS-CULTURE
REGULATION OF THE ALGINATE BIOSYNTHESIS GENE ALGC IN PSEUDOMONAS-AERUGINOSA DURING BIOFILM DEVELOPMENT IN CONTINUOUS-CULTURE
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DOI：
10.1128/aem.61.3.860-867.1995
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发表时间：
1995-03-01
影响因子：
4.4
通讯作者：
GEESEY, GG
中科院分区：
文献类型：
--
作者：
DAVIES, DG;GEESEY, GG
采用报告基因技术观察铜绿假单胞菌粘液菌株中藻酸盐生物合成基因algC在聚四氟乙烯和玻璃基质上连续培养中发育和成熟生物膜的调控情况，携带algC-lacZ转录融合体的质粒pNZ63在无选择压力的情况下连续培养25天不被稀释，本体相稳态条件下的生物膜细胞表现出波动16 天期间 algC 表达的变化，但没有表明随时间间隔表达增加或减少的趋势。通过使用显微镜结合图像分析，使用质粒携带的 lacZ 基因产物（β-半乳糖苷酶）的荧光底物对发育中的生物膜中 algC 上表达进行了体内检测。通过这种技术，随着时间的推移跟踪细胞并分析 algC 活性。在生物膜形成的初始阶段，已经附着于玻璃表面至少15分钟的细胞表现出algC的上表达，可通过全细胞荧光的发展来检测。然而，最初的细胞附着于基质似乎与algC启动子活性无关。此外，未表现出algC上表达的细胞在流动条件下表现出比检测到algC上表达的细胞在流动条件下保留在玻璃表面的能力更弱。
Reporter gene technology was used to observe the regulation of the alginate biosynthesis gene, algC in a mucoid strain of Pseudomonas aeruginosa in developing and mature biofilms in continuous culture on Teflon and glass substrata, The plasmid pNZ63, carrying an algC-lacZ transcriptional fusion, was shown to not be diluted in continuous culture over a period of 25 days in the absence of selection pressure, Biofilm cells under bulk phase steady-state conditions demonstrated fluctuations in algC expression over a 16-day period, but no trend of increased or decreased expression over the time interval was indicated, In vivo detection of algC up-expression in developing biofilms was performed with a fluorogenic substrate for the plasmid-borne lacZ gene product (beta-galactosidase) by using microscopy coupled with image analysis, By this technique, cells were tracked over time and analyzed for algC activity. During the initial stages of biofilm development, cells already attached to a glass surface for at least 15 min exhibited up-expression of algC, detectable as the development of whole-cell fluorescence, However, initial cell attachment to the substratum appeared to be independent of algC promoter activity, Furthermore, cells not exhibiting algC up-expression were shown to be less capable of remaining at a glass surface under flowing conditions than were cells in which algC up expression was detected.