耐辐射Belnapia菌株的分离和辐射后蛋白质组学分析  

Identification of a Radiation Resistant Belnapia Strain and Proteomic Analysis after Exposure to Gamma Radiation

作　　者：白明艳[1] 宣慧娟[1] 李利[1] 孙云峰[1] 万平[1] 杨志伟[1] 

机构地区：[1]首都师范大学生命科学学院,北京100048

出　　处：《核农学报》2014年第2期169-176,共8页Journal of Nuclear Agricultural Sciences

基　　金：北京市属高等学校人才强教深化计划-极端环境植物微生物资源挖掘(2010)

摘　　要：从西藏土样中分离得到一株具有较强辐射抗性的革兰氏阴性菌株F-4,其16S rDNA序列与Belnapia属菌株具有很高的相似性。结合表型特点,初步将其鉴定为别尔纳普氏属(Belnapia)菌株。在UV和γ-射线辐照后,菌株F-4的D10分别为300J·m-2和4000Gy,远高于辐射敏感的E.coli K12。为研究菌株F-4在蛋白质组水平对辐射的响应机制,将对数早期的F-4细胞置于1000Gyγ-射线辐照下,并在5h复苏培养后,对其可溶性蛋白进行了提取、双向电泳分析和MALDI-TOF/TOF质谱鉴定。结果表明,辐射后32个蛋白质的表达出现3倍以上的变化,其中24个蛋白质上调,8个蛋白质下调,涉及蛋白质折叠加工、能量代谢、氨基酸代谢、辅酶代谢和一些功能未知途径。本试验为研究细菌辐射抗性机制提供了一些启示。A radiation resistant gram-negative Bacterium strain, F -4, was isolated from the soil samples collected from the central Tibet. The 16S rDNA sequence of F - 4 has high similarities with those species in genus Belnapia. Combined with morphology characteristics, strain F- 4 was proposed the name Belnapia sp. F -4. The D^0 values of F -4 after exposure to UV or gamma radiation were 300J-m-2 and 4000Gy respectively, much higher than those of radio-sensitive E. coli K12. The proteomic changes in response to gamma radiation were analyzed by exposing the early exponential F - 4 cells to 1000 Gy gamma rays at room temperature. After irradiation, the ceils was cultivated for 5 hours for recovery, and then the cellular soluble proteins were extracted and seperated by two-dimensional electrophoresis. MALDI-TOF-MS analysis was carried out to identify the differential expressed proteins after gamma irradiation. The results showed that 32 proteins displayed 3 - fold increase or decrease after irradiation. Among them, 24 proteins were up-regulated and 8 proteins were down-regulated. These responsive proteins participate in protein folding, energy metabolism, amino acid metabolism and unknown pathways. Studies on the radio-resistant Belnapia sp. F -4 may cast some light on elucidating the mechanism of radiation tolerance in bacteria.

关 键 词：辐射抗性 别尔纳普氏菌 耐辐射奇异球菌 蛋白质组学分析 

分 类 号：Q936[生物学—微生物学]