蠕形螨全蛋白提取及相对分子量鉴定  

Extraction of total proteins from demodex and qualification of their molecular weights

作　　者：朱晓丽[1] 郭淑玲[1] 苏磊[1] 冯玉新[2] 袁方曙[1] 

机构地区：[1]山东大学医学院病原生物学研究所,山东济南250012 [2]山东大学医学院细胞生物学研究所,山东济南250012

出　　处：《山东大学学报（医学版）》2014年第5期58-62,共5页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(Y2008C111)

摘　　要：目的探讨蠕形螨全蛋白提取方法,初步分析蠕形螨蛋白组分。方法采用自制裂解液和RIPA裂解液分别提取蠕形螨全蛋白,通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分离比较蛋白提取效果,用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)鉴定部分蛋白质条带,Mascot检索软件查询NCBInr数据库,对蠕形螨蛋白组分进行初步分析。结果自制裂解液提取蠕形螨蛋白的效果优于RIPA裂解液。经数据库检索比对出21种可信蛋白,其中170 kD条带比对出的蛋白为肌动蛋白和胞质肌动蛋白,130 kD条带比对出的蛋白为谷氨酰胺连接酶和腺苷酰转移酶;15 kD以下条带比对出的蛋白大部分为组蛋白。结论自制裂解液液氮研磨法可提取到较高质量的蠕形螨全蛋白,实验提取并分析了蠕形螨的蛋白质组成,为蠕形螨蛋白组学研究奠定基础。Objective To establish the optimal extraction method of demodex protein and to analyze its components. Methods Demodex proteins were extracted by self-made lysate and RIPA lysate,and the extraction effects were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE）. Partial protein stripes were identified with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF-MS）. The NCBInr database was searched by Mascot software to analyze the components of demodex protein. Results The extraction effect of self-made lysate was better than that of RIPA protein lysate. A total of 21 proteins were primarily identified. Conclusion The protein components of demodex were extracted in our experiment,which is helpful for the future study on demodex proteomics.

关 键 词：蠕形螨 十二烷基硫酸钠聚丙烯酰胺凝胶电泳 蛋白质组学 分子量 

分 类 号：R384.4[医药卫生—医学寄生虫学]