裂解液对变异链球菌蛋白质提取效果的影响

Influence of Cell Lysis Buffers on the Effect of Protein Extraction from Streptococcus Mutans

目的探讨裂解液对变异链球菌蛋白质提取效果的影响。方法通过革兰染色、SDS-PAGE和蛋白质浓度测定,对4种裂解液提取变异链球菌蛋白质的效果进行评价。结果裂解液Ⅲ、Ⅳ中的细菌破碎程度高于裂解液Ⅰ、Ⅱ。裂解液Ⅱ提取的蛋白质条带丰度低于其他3种裂解液。裂解液Ⅲ、Ⅳ提取的蛋白浓度高于裂解液Ⅰ、Ⅱ(P<0.05)。结论含还原剂、去垢剂、离液剂和蛋白酶抑制剂的裂解液提取变异链球菌蛋白质的效果优于只含还原剂和去垢剂的裂解液。

Objective To investigate the influence of different cell lysis buffers on the effect of extracting protein from Streptococcus . mutans（S.mutans）. Methods Four kinds of cell lysis buffers containing different constituents were used to extract proteins from S.mutans. Their effects were evaluated through Gram＇s staining of bacterial suspension smear, SDS-PAGE of protein and measurement of extracted proteins＇ concentration. Results The breaking degree of S.mutans treated in cell lysis buffer m and IV was higher than in cell lysis buffer Ⅰ and lI. The abundance of the extracted proteins＇ strap in cell lysis buffer Ⅱ was obviously lower than in other three cell lysis buffers. The concentration of the extracted proteins in cell lysis buffer Ⅲ and Ⅳ was significantly higher than that in cell lysis buffer Ⅰ and Ⅱ （P〈0.05）. Conclusion Cell lysis buffers simuhaneously containing reducing agent, detergent, chaotropic agent and proteinase inhibitor had obviously better effect of extracting proteins from S.mutans, compared with that only containing reducing agent and detergent.