Starck Joakim, Källenius Gunilla, Marklund Britt-Inger, Andersson Dan I, Åkerlund Thomas
Department of Bacteriology, Swedish Institute for Infectious Disease Control, S-17182, Solna, Sweden.
Microbiology and Tumour Biology Centre, Karolinska Institute, S-17177, Stockholm, Sweden.
Microbiology (Reading). 2004 Nov;150(Pt 11):3821-3829. doi: 10.1099/mic.0.27284-0.
Data are presented from two-dimensional (2-D) PAGE analysis of Mycobacterium tuberculosis strain Harlingen grown during aerobic and anaerobic culture, according to a modified Wayne dormancy model. M. tuberculosis cultures were grown to the transition point between exponential growth and stationary phase in the presence of oxygen (7 days) and then part of the cultures was shifted to anaerobic conditions for 16 days. Growth declined similarly during aerobic and anaerobic conditions, whereas the ATP consumption rapidly decreased in the anaerobic cultures. 2-D PAGE revealed 50 protein spots that were either unique to, or more abundant during, anaerobic conditions and 16 of these were identified by MALDI-TOF. These proteins were the alpha-crystalline homologue (HspX), elongation factor Tu (Tuf), GroEL2, succinyl-CoA : 3-oxoacid-CoA transferase (ScoB), mycolic acid synthase (CmaA2), thioredoxin (TrxB2), beta-ketoacyl-ACP synthase (KasB), l-alanine dehydrogenase (Ald), Rv2005c, Rv2629, Rv0560c, Rv2185c and Rv3866. Some protein spots were found to be proteolytic fragments, e.g. HspX and GroEL2. These data suggest that M. tuberculosis induces expression of about 1 % of its genes in response to dormancy.
根据改良的韦恩休眠模型，展示了在需氧和厌氧培养条件下生长的结核分枝杆菌哈林根菌株的二维（2-D）聚丙烯酰胺凝胶电泳分析数据。结核分枝杆菌培养物在有氧条件下（7天）生长至指数生长期和稳定期之间的过渡点，然后将部分培养物转移至厌氧条件下培养16天。在需氧和厌氧条件下生长均同样下降，而厌氧培养物中的ATP消耗迅速减少。二维聚丙烯酰胺凝胶电泳显示有50个蛋白点在厌氧条件下是独特的或更为丰富，其中16个通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）进行了鉴定。这些蛋白质包括α-晶体同源物（HspX）、延伸因子Tu（Tuf）、GroEL2、琥珀酰辅酶A：3-氧代酸辅酶A转移酶（ScoB）、分枝菌酸合酶（CmaA2）、硫氧还蛋白（TrxB2）、β-酮脂酰-ACP合酶（KasB）、L-丙氨酸脱氢酶（Ald）、Rv2005c、Rv2629、Rv0560c、Rv2185c和Rv3866。发现一些蛋白点是蛋白水解片段，例如HspX和GroEL2。这些数据表明结核分枝杆菌在休眠时诱导约1%的基因表达。
