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[Cloning of phzIR from the endophytic Pseudomonas sp. G5 and its expression in Escherichia coli].
- Source :
-
Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2009 Jun; Vol. 25 (6), pp. 832-9. - Publication Year :
- 2009
-
Abstract
- We isolated a new strain of endophytic Pseudomonas G5 from the stems of Chinese parsley (Coriandrum sativum L.), and it is tentatively identified as Pseudomonas aurantiaca according to analysis of the entire substrate utilization profiles using BIOLOG Microstation system (BIOLOG, Inc, Hayward CA). An array of evidence established that many Gram-negative bacteria employ Quorum sensing (QS) system to regulate gene expression in response to cell density using small diffusible signal molecules, N-acyl homoserine lactones (AHLs), and control diverse phenotypic traits in plant-associated bacteria. In this study, we showed that Pseudomonas sp. strain G5 can produce several types of AHLs at a detectable level using Thin Layer Chromatography (TLC) analysis combined with bioreporter Chromobacterium violaceum CV026 bioassay, and N-hexanoyl-homoserine lactone (HHL, C6-HSL) with Rf value 0.4 is the major signal molecule. Furthermore, we have identified its quorum sensing system composed of PhzI and PhzR by cloning and sequencing of phzI-phzR. PhzI is responsible for synthesis of AHLs signal molecules, and PhzR is a transcriptional regulator. Finally, we heterologously expressed the recombinant plasmid pMD-phzIR in Escherichia coli JM109 and verified it using C. violaceum CV026 bioassay. The phylogenetic analysis using MEGA4 revealed highly similarities exist among the phzIR homologs, suggesting it is evolutionary well conserved in the genus Pseudomonas.
- Subjects :
- 4-Butyrolactone analogs & derivatives
4-Butyrolactone metabolism
Acyl-Butyrolactones metabolism
Amino Acid Sequence
Bacterial Proteins biosynthesis
Cloning, Molecular
Escherichia coli genetics
Gene Expression Regulation, Bacterial
Genetic Vectors genetics
Molecular Sequence Data
Phylogeny
Pseudomonas classification
Pseudomonas isolation & purification
Trans-Activators biosynthesis
Bacterial Proteins genetics
Escherichia coli metabolism
Pseudomonas genetics
Trans-Activators genetics
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-3061
- Volume :
- 25
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 19777809