家畜生态学报2018，Vol.39Issue(3)：12-16,5.
枯草芽孢杆菌突变株的构建及其对猪肺泡巨噬细胞(MΦ3D4/2)免疫因子表达的影响
Construction of Bacillus Subtilis Mutant Strain and its Effect on Immune Factor Expression of Porcine Alveolar Macrophages(MΦ3D4/2)
摘要
Abstract
To examine the ability of Bacillus.Subtilis spores and vegetative cells to affect the secretion of porcine alveolar macrophages cytokines,we first knocked out the spore germination gene clW-gerD and sporulation gene spo0A from chromosomal DNA of B.subtilis PY79,constructed the deletion mutants B.subtilis clW-gerD-and B.subtilis spo0A-,extracted genomic DNA for PCR detection.The spore germina tion ability of spore germination mutant strains and the sporulation ability of spore mutant strain were evaluated by Gram staining and germination rate detection.The secretion expression level of cytokines was detected by ELISA test kit after B.subtilis clW-gerD-and B.subtilis spo0A-respectively cocultured of the porcine alveolar macrophages.We successfully deleted the spore germination gene clW-gerD-and sporulation gene spo0A,constructed the spore germination mutants B.subtilis clW-gerD-and the sporulation mutant B.subtilis spo0A-.The results of the sporulation ability and sporulation ability showed that B.subtilis clW-gerD-of three spore germination mutants completely deprived spore germination ability.Microscope observation and spore production rate test indicated that B.subtilis spo0A-completely lost sporulation ability.In coculture experiment,B.subtilis clW-gerD-significantly increased the expression of cytokine IL-6(P＜0.01);Whereas the expression of IL-1β,IL-10 and IL-8 showed a less pronounced increase after B.subtilis clW-gerD-,B.subtilis PY7 and B.subtilis spo0A-interacted with macrophage (P ＞0.05).In summary,B.subtilis clW-gerD-could stimulate the macrophage secretion of proinflammatory factor IL-6,strengthen inflammatory reaction of MΦ3D4/2 cells and improve the ability of anti-infection.关键词
枯草芽孢杆菌/猪肺泡巨噬细胞/免疫功能/细胞因子Key words
Bacillus Subtilis/porcine alveolar macrophages/immunity/cytokine分类
农业科技引用本文复制引用
李天瑾,杨明明,龚月生..枯草芽孢杆菌突变株的构建及其对猪肺泡巨噬细胞(MΦ3D4/2)免疫因子表达的影响[J].家畜生态学报,2018,39(3):12-16,5.基金项目
国家自然科学基金(31372343) （31372343）
