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目的研究回转器模拟微重力对革兰氏阴性菌大肠杆菌(Escherichia coli)基因与蛋白表达的影响。方法采用回转器模拟微重力效应,连续2周作用于大肠杆菌ATCC 25922菌株,选取16个靶基因,RT-PCR反应检测处理菌株和对照菌株基因表达的变化;提取菌体蛋白进行双向电泳,电泳图谱采用ImageMaster 2D Platinum软件分析,找出差异表达蛋白,进行基质辅助激光解吸飞行时间质谱(MALDI TOF/TOF质谱)分析。结果在模拟微重力效应作用下,处理菌株与对照菌株相比,16个靶基因中有4个靶基因表达上调,5个下调;15个差异表达蛋白点,其中4个表达上调蛋白质谱鉴定为抗转录终止蛋白NusG、固氮铁蛋白、硫醇过氧化物酶,1个表达下调蛋白为未知蛋白,2个新增蛋白为谷氨酰胺ABC转运周质蛋白,1个表达消失蛋白为IclR转录调节蛋白家族。结论模拟微重力效应可引起大肠杆菌基因及功能蛋白表达的变化。
Objective To study the effects of rotameter simulated microgravity on the gene and protein expression of gram-negative Escherichia coli. Methods Rotavators were used to simulate the effects of microgravity and were applied to Escherichia coli ATCC 25922 for 2 consecutive weeks. Sixteen target genes were selected and the changes of gene expression of the treated and control strains were detected by RT-PCR. Two-dimensional gel electrophoresis The maps were analyzed with ImageMaster 2D Platinum software to identify differentially expressed proteins for MALDI-TOF / TOF mass spectrometry analysis. Results Under simulated microgravity effect, compared with the control strain, 4 of the 16 target genes were up-regulated and 5 down-regulated compared with the control strain. Fifteen differentially expressed protein spots, of which four up-regulated proteins were identified as Transcription termination protein NusG, ferritin, thiol peroxidase, an expression of down-regulated protein unknown protein, two new protein glutamine ABC transporter periplasmic proteins, one expression of the disappearance of protein IclR transcriptional regulation Protein family. Conclusion Simulated microgravity effect can cause the changes of E. coli gene and function protein expression.