摘要(Abstract)：
为了获得托高土病毒核蛋白氮端结构域的蛋白(THOV-NP_N)晶体，进一步探究其结构和功能，依次采用镍亲和层析、肝素亲和层析方法纯化大肠杆菌异源表达获得的THOV-NP_N,利用凝胶过滤层析及十二烷基磺酸钠-聚丙烯酰胺凝胶电泳法检测THOV-NP_N的均一性和纯度，分别用坐滴法和悬滴法筛选和优化THOV-NP_N结晶条件，采用质谱法验证所得的晶体。结果表明：所使用的提取方法可以获得纯度较高、性质均一的THOV-NP_N,在培养温度16℃, THOV-NP_N的质量浓度为25.0 g/L,池液中磷酸氢二铵、柠檬酸钠、氯化钠的浓度分别为0.8、 0.1、 0.2 mol/L及pH=5.0的条件下可以获得粒径为0.05 mm、形状近似正方体的THOV-NP_N晶体。
关键词(KeyWords)： 托高土病毒;核蛋白;异源表达;纯化;结晶
基金项目(Foundation): 山东省自然科学基金项目(ZR2020QC057);; 泰山学者青年专家计划项目(tsqn201812079)
作者(Author): 马林,鞠金鑫,李琳,秦晓春,董士尚
DOI: 10.13349/j.cnki.jdxbn.20240308.001
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