结核分枝杆菌生物膜形成相关基因的筛选与鉴定-遗传学专业论文.docxVIP

宁夏医科大学硕士研究生学位论文中文摘要 宁夏医科大学硕士研究生学位论文 中文摘要 II II 物膜形成调控机制提供候选基因，为后续深入开展生物膜形成机制研究奠定基础。 关键词 结核分枝杆菌，MycoMarT7 噬菌体转座子，菌落形态，生物膜 宁夏医科大学硕士研究生学位论文英文摘要 宁夏医科大学硕士研究生学位论文 英文摘要 III III Screening and Identification of Mycobacterium tuberculosis Biofilm Formation Related Genes 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. Compared with the wild-type strain, the ability to form biofilms which can be formed biofilm in the 20 ~ 25d of 8 strains has not occurred any change in the biofilm. The P7C8 mutant was the biofilm completely deficient strain, there were still no biofilm formation phenotype which observed after cultured 7 weeks. The remaining 7 mutant had the phenomenon of biofilm growth or maturation delayed, one of the two mutant formed biofilms had taken up to 30 days, 5 strains of about had began to be formed the thin film layer which can clearly be seen by the naked eye in the surface of the gas-liquid after 40 days. 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 v