肺炎链球菌SP0306蛋白的表达纯化及结晶研究

doi:10.13523/j.cb.20150604

中国生物工程杂志

2015, Vol. 35

Issue (6): 21-25 DOI: 10.13523/j.cb.20150604

研究报告

肺炎链球菌SP0306蛋白的表达纯化及结晶研究

黄健¹, 黄美容², 朱杰华¹, 骆诗露¹, 闵迅¹

1. 遵义医学院附属医院检验科遵义 563003;
2. 遵义医学院附属医院输血科遵义 563003

Expression,Purification,Crystallization of SP0306 Protein from Streptococcus pneumoniae

HUANG Jian¹, HUANG Mei-rong², ZHU Jie-hua¹, LUO Shi-lu¹, MIN Xun¹

1. Affiliated Hospital of Zunyi Medical College, Department of Laboratory Medicine, Zunyi 563003, China;
2. Affiliated Hospital of Zunyi Medical College, Department of Blood Transfusion, Zunyi 563003, China

全文: PDF(778 KB) HTML

摘要：

SP0306蛋白是肺炎链球菌TIGR4菌株中的一种假想的转录因子,但其蛋白三维结构及生物学功能尚未明了,生物信息学分析提示其可能调控碳水化合物代谢相关基因的表达。成功构建了SP0306蛋白的全长表达载体PET28a-sp0306,利用大肠杆菌BL21(DE3)菌株进行原核表达,获得了以可溶形式表达的目的蛋白。经Ni-NTA柱亲和层析及DEAE阴性离子交换层析纯化后,获得了高纯度的目的蛋白。采用悬滴气相扩散法获得了质量较好的SP0306蛋白晶体,并初步进行了晶体X射线衍射,为其最终的三维结构解析及生物学功能研究奠定了基础。

关键词： 肺炎链球菌; SP0306蛋白; 原核表达; 晶体生长

Abstract:

SP0306 protein is a hypothetical transcription factor of Streptococcus pneumoniae TIGR4 strains, but the protein three-dimensional structure and biological function is unclear. The bioinformatics analyses show that it may regulate the expression of carbohydrate metabolism related genes. The protein expression vector of PET28a-sp0306 was successfully constructed and a soluble form of SP0306 protein was acquired in Escherichia coli BL21 (DE3) strain. High purity SP0306 protein was obtained after Ni-NTA affinity chromatography and DEAE negative ion exchange chromatography. High quality crystal of SP0306 protein was obtained by the vapor diffusion method, then crystal X-ray diffraction was executed.The results provide a basis for helping resolve the structure of SP0306 and further function research.

Key words: Streptococcus pneumoniae SP0306 protein Prokaryotic expression Crystal grow

收稿日期: 2015-03-05 出版日期: 2015-06-25

ZTFLH:

Q819

基金资助:

国家自然科学基金(81460317)、贵州省社会发展公关计划(2012GZ83293)资助项目

通讯作者: 闵迅 E-mail: zmchj2001@163.com

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引用本文:

黄健, 黄美容, 朱杰华, 骆诗露, 闵迅. 肺炎链球菌SP0306蛋白的表达纯化及结晶研究[J]. 中国生物工程杂志, 2015, 35(6): 21-25.

HUANG Jian, HUANG Mei-rong, ZHU Jie-hua, LUO Shi-lu, MIN Xun. Expression,Purification,Crystallization of SP0306 Protein from Streptococcus pneumoniae. China Biotechnology, 2015, 35(6): 21-25.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150604 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I6/21

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