非分型流感嗜血杆菌Haps蛋白表达量影响因素的探索  

Impacting factors for Haps protein expression in non-typeable Haemophilus influenzae

作　　者：邝玉[1] 李婉宜[1] 李明远[1] 李虹[1] 曾蔚[1] 马巨辉[1] 

机构地区：[1]四川大学华西基础医学与法医学院微生物学教研室,四川成都610041

出　　处：《西部医学》2006年第1期7-9,共3页Medical Journal of West China

摘　　要：目的优化非分型流感嗜血杆菌HapS蛋白质片段表达与分泌量的条件。方法用BHI液体培养基培养非分型流感嗜血杆菌(NTHi),观察不同培养时间对NTHi培养液光吸收值的影响和HapS蛋白质片段分泌量的变化;NTHi在BHI培养液中培养18 h后,加入IPTG诱导不同时间,观察其对HapS蛋白质片段分泌量的影响。结果增加NTHi在BHI中的培养时间,培养液的光吸收值呈下降趋势,其培养上清提取的蛋白质样品经SDS-PAGE电泳没有显示出HapS蛋白质片段(110KD)的条带;加入IPTG诱导80min后,提取的蛋白质样品经SDS-PAGE电泳后,显示出有明显的110KD的条带。结论增加培养时间不能增加NTHi的细菌总数和HapS蛋白质片段的表达与分泌量,而IPTG能诱导NTHi细菌hap基因的表达,明显增加HapS蛋白质片段的分泌量。Objective To explore the good conditions for the secretion of Haps protein of non-typeable Haemophilus influenzae （NTHi）. Methods Prolong the NTHi culture time in BHI medium, and then observe the changes of absorbency of cultured liquid at 600 micrometer （A600） and secretion of Haps in the supernatants. We also added IPTG to the culture medium to observe the inducing expression of the Haps gene. Results The A600 yalue of NTHi culture medium decreased continually after prolonged the cultured time in BHI medium, and the Haps band （110KD） was not shown obviously when protein saraple extracted from the bacterial culture supernatants was analyzed by SDS-PAGE. After the NTHi was induced by IPTG for 80min, there were obvious 110KD bands in the protein sample extracted from the NTHi culture supernatant with SDS-PAGE. Conclusion The amount of the bacterial and the secretion of Haps protein do not increase through extended cultured time. IPTG could induce the expression of the hap gene, and increase the secretion of HapS protein.

关 键 词：非分型流感嗜血杆菌 HAPS 培养时间 IPTG 白表达量 

分 类 号：R378.4[医药卫生—病原生物学]