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| Screening and Identification of Mycobacterium tuberculosis Biofilm Formation Related Genes |
| LIU Xia1,2, GUO Qing-long1, WANG Ruo-jun1, WANG Hong-hai1, PEI Xiu-ying2, ZHANG Xue-lian1 |
1. State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China;
2. Biochemistry and Molecular Biology Department of School of Basic Medical Sciences, Ningxia Medical University Key Laboratory of Reproduction and Heredity of Ningxia Hui Autonomous Region, Medical Sci-Tech Research Center, Yinchuan 750004, China |
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Abstract Objective:To screen and identify the related genes which may be associated with biofilm formation of Mycobacterium tuberculosis (M. tuberculosis) based on the changes of colony phenotype and biofilm defects. Methods:A random mutagenesis library was constructed by using the MycoMarT7 transposon system. Mutants with colony morphological changes on solid agar plate or with defects in pellicle biofilm formation in liquid culture medium were selected. In order to determine the locus of mutation gene, the flanking fragments of gene were obtained using T-A cloning and resistance marker recovery method. Then, the function of mutated genes was analyzed and predicted by bioinformatics methods. Results: 39 mutants with colony morphological changes or biofilm defects were selected and the loci interrupted of 16 mutants were successfully identified. Among the 16 genes, 5 genes were associated with the regulation of lipid metabolism, 4 were participated in cell wall and cell processes, 2 were related to intermediary metabolism and respiration, the product of 1 gene might be a regulatory protein, 1 was reported as virulence gene in previous studies, 1 was PE/PPE family gene and the function of other genes were unknown. Among these genes, 8 genes might be related to deficient in forming pellicle biofilm. Conclusions: A M. tuberculosis transposon insertion mutant library of approximately 10 000 mutants was successfully constructed, pellicle biofilm formation defects were selected and biofilm formation related genes were identified. All results will be the basis of the further research of biofilm formation mechanism in M. tuberculosis.
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Received: 21 January 2013
Published: 25 April 2013
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