荧光假单胞菌中蛋白OpuAC的初步晶体学和生物学功能的研究
preliminary study on crystallography and biochemical function of protein OpuAC from pseudomonas fluorescens
目的：探究荧光假单胞菌中ATP结合盒式蛋白OpuAC的晶体三维结构和生化功能。方法：用PCR方法获取编码OpuAC的基因片段，构建重组表达载体(pET-28b-OpuAC)并克隆至大肠杆菌表达菌株BL21（DE3）中，然后通过异丙基-β- D-硫代半乳糖苷诱导表达，DEAE-Sepharose Fast Flow(SPFF)纯化，浓缩OpuAC（>10 mg/ml）重组蛋白，同时用thermal shift 的方法探究蛋白OpuAC对渗透性相容物质的调节作用。结果: 成功获得了高纯度的OpuAC蛋白，并得到了OpuAC蛋白晶体，同时研究得到蛋白OpuAC能够较强的结合渗透性相容物质-甜菜碱。结论：得到OpuAC 蛋白晶体和其与甜菜碱的结合为进一步解析三维结构和生化分子机制的研究奠定了基础。?????
Objective To explore the three-dimensional structure and biochemical function of ATP-binding cassette protein OpuAC from pseudomonas fluorescens. Methods The OpuAC gene was amplified by PCR, cloned into the plasmid pET-28b and then the recombinant plasmid was transformed into E.coli BL21(DE3) cells. The protein of OpuAC was obtained by DEAE-Sepharose Fast Flow after inducing by isopropyl β-D-thiogalactoside. Meanwhile, the ability of OpuAC mediating the compatible solutes was studied by Protein Thermal Shift Assay. Result The crystals of OpuAC with high concentrations were successfully obtained. In addition, the protein was tightly combined with glycine betaine. Conclusion These results may contribute to the further research on the 3-D structure and biochemical molecular mechanism of OpuAC from pseudomonas fluorescens .?????
何永兴、郭玎玎、韩建庭、张孟元、张四萍、王勇
生物化学分子生物学微生物学
OpuAC蛋白原核表达晶体筛选甜菜碱热能量转移。
Protein OpuACprokaryotic expressioncrystal screeningthermal shiftglycine betaine.?????
何永兴,郭玎玎,韩建庭,张孟元,张四萍,王勇.荧光假单胞菌中蛋白OpuAC的初步晶体学和生物学功能的研究[EB/OL].(2017-12-08)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/201712-135.点此复制
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