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Author:
Date:
2013-07-05
[Abstract] This protocol was developed in a study aimed to determine the cellular localization of the lysin of pneumococcal phage SV1 (Frias et al., 2013). We obtained proteins from the surface of Streptococcus pneumoniae by elution with choline or those secreted to the medium. The analysis by Western blot of these fractions allowed us to demonstrate that the phage lysin localizes to the cell wall, associating with choline residues in the teichoic acids. Hence, protein extracts can be used to determine the localization of uncharacterized proteins and can also be useful for other biochemical analyses such as protein identification. This protocol can be easily adapted to different pneumococcal strains and growth conditions and it is well suited to isolate other proteins of interest.
[摘要] 该方案是在旨在确定肺炎球菌噬菌体SV1的溶素的细胞定位的研究中开发的（Frias等人，2013）。 我们通过用胆碱或分泌到培养基的那些洗脱从肺炎链球菌的表面获得蛋白质。 通过这些级分的Western印迹分析允许我们证明噬菌体溶素定位于细胞壁，与磷壁酸中的胆碱残基相关。 因此，蛋白质提取物可用于确定未表征蛋白质的定位，并且还可用于其他生物化学分析，例如蛋白质鉴定。 该方案可以容易地适应于不同的肺炎球菌菌株和生长条件，并且其非常适合于分离其他感兴趣的蛋白质。
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