耻垢分枝杆菌7天培养滤液与Middlebrook 7H9培养液蛋白质组对比分析  

COMPARATIVE ANALYSIS OF PROTEOME BETWEEN MEDIUM LIQUID OF MIDDLEBROOK 7H9 AND 7-DAY CULTURE FILTRATE OF THE M.SMEGMATIS

作　　者：张为民[1,2] 吴劲松[1,2] 龚文波[1,2] 

机构地区：[1]深圳市人民医院 [2]暨南大学第二临床学院检验医学部,广东深圳518020

出　　处：《现代医院》2013年第8期8-10,共3页Modern Hospitals

摘　　要：目的初步了解耻垢分枝杆菌体外培养过程中对蛋白质的利用及其蛋白分泌活动状况，以进一步了解其生物特性。方法应用与弱阳离子交换蛋白质芯片（WCX2）联用的表面增强激光解吸一电离飞行时间质谱（SELDI—TOF—MS）技术检测Middlebrook7H9培养液和耻垢分枝杆菌7天培养滤液蛋白质／多肽，采用描述性分析其蛋白质／多肽谱并比较两者间的差异。结果在Middlebrook7H9培养液中检测到97种蛋白质／多肽，相对分子质量在1050～3940之间，丰度水平在86～7838相对强度值之间（相对强度值通过质谱峰面积计算）；在耻垢分枝杆菌7天培养滤液中检测到125种蛋白质／多肽，相对分子质量在1050～9330之间，丰度水平在111～9961相对强度值之间；相对Middlebrook7H9培养液，耻垢分枝杆菌7天培养滤液的蛋白质／多肽谱发生了下列变化：45种消失、20种丰度降低、3种未变化、30种丰度增大、71种新产生。结论耻垢分枝杆菌作为快速生长分枝杆菌菌种，在体外培养生长时有活跃的蛋白质利用和分泌活动，对于既无侵袭力又无毒素产生的耻垢分枝杆菌而言，这些活动可能与其致病性密切相关，值得人们深入研究。Objective To study primarily the information on use of protein in medium and protein secretion in the course of vitro culture of M. smegmatis and understand the biological character of this germ. Methods Protein/peptides of medium liquid of Middlebrook 7H9 and 7 - day culture filtrate of the M. smegmatis captured with weak cation exchange pro-tein chip（ WCX2 bead） were detected by surface - enhanced laser desorption/ionization time - of - flight mass spectrometry （ SELDI - TOF - MS）. Descriptive analysis and comparison were performed. Resdts 97 kinds of protein/peptides whose rel-ative molecular masses range from 1 050 to 3 940 and levels of abundance with relative intensity value calculated by mass spectrometry peak area were in the range of 86 to 7 838 were detected in the medium liquid of Middlebrook 7H9 while they are 125, from 1 050 to 9 330 and from 111 to 9 961 in the 7 - day culture filtrate of the M. smegmatis. Compared with the medi- um liquid of Middlebrook 7H9, the style was present such as disappearing of 45 p kinds of protein/peptides, decrease of a- bundance of 20 kinds of protein/peptides, increase of abundance of 30 kinds of protein/peptides, 71 kinds of new protein/ peptides and no change of 3 kinds of protein/peptides. Conclusion There are strong protein utilization and secretory activity in the course of vitro culture of M. smegmatis belonged to a fast - growing mycobacterium species. Toward M. Smegmatis as a sort of without invasion ability and toxin creation mycobacteria, these activities may be intimate correlate with its pathogenic-ity and worthy of study thoroughly.

关 键 词：耻垢分枝杆菌 培养滤液 蛋白质组 弱阳离子交换蛋白质芯片(WCX2) 表面增强激光解吸-电离飞行时间质谱(SELDI—TOF—MS) 

分 类 号：R378.911[医药卫生—病原生物学]