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目的:对空肠弯曲菌(Cj)的双向电泳(2DE)系统可重复性和稳定性进行评价,为空肠弯曲菌的蛋白质组学研究奠定基础。方法:选取一株空肠弯曲菌,培养48小时后制备全菌蛋白,进行双向电泳。比较不同上样量和不同批次对实验结果的影响。结果:不同批次和不同上样量的胶进行比较,蛋白质的匹配率均可达到90.89%;未匹配蛋白主要是一些低丰度蛋白;增加上样量可增加低丰度蛋白的检出,但可产生蛋白质的共迁移,因此选择合适的上样量很重要;不同批次蛋白质点在等电点方向的位置偏移为0.62±0.37mm,在SDS-PAGE方向的位置偏差为0.72±0.51mm,相对含量的相关系数为0.973。结论:表明空肠弯曲菌2DE的系统具有很好的稳定性及很高的可重复性,符合蛋白质组学的要求,实验结果具有可比性。
OBJECTIVE: To evaluate the repeatability and stability of two-dimensional electrophoresis (2DE) system of Campylobacter jejuni (Cj) and lay a foundation for the proteomic study of Campylobacter jejuni. Methods: One strain of Campylobacter jejuni was selected and the whole bacterial protein was prepared after 48 hours of cultivation. The two-dimensional electrophoresis was performed. The effects of different loadings and batches on the experimental results were compared. Results: Compared with different loadings of different batches of samples, the match rate of the proteins reached 90.89%. The unmatched proteins were mainly low-abundance proteins. Increasing the loading amount could increase the detection of low-abundance proteins, But can produce co-migration of proteins, so it is very important to choose the appropriate sample loading capacity. The protein spots in different batches have a position shift of 0.62 ± 0.37mm in the isoelectric point and a position deviation of 0.72 ± 0.51 in the SDS-PAGE mm, the relative content of the correlation coefficient of 0.973. Conclusion: The results show that the 2DE system of Campylobacter jejuni has good stability and reproducibility, which meets the requirements of proteomics. The experimental results are comparable.