摘要
目的:优化重组人白细胞介素17的原核表达条件,选择不同的路线进行分离纯化.方法:利用SDS-PAGE方法分析比较重组人白细胞介素17在不同培养基、不同诱导时间的表达量,选择最适条件,大量扩增后,分离出包涵体,分别采用盐酸胍溶解法和SDS溶解法两条纯化路线进行进一步的分离和纯化,并对所纯化产物进行常规分析鉴定.结果:采用上述两种不同的溶解包涵体方法,通过后续复性及纯化,均可得到高纯度具有生物学活性的重组IL-17.采用SDS溶解法具有较高的得率,采用盐酸胍溶解法纯化得率较低,得产物的生物学活性较高.结论:通过表达及纯化,得到了大量高纯度具有活性的rhIL-17,为进一步研究IL-17的体内外生物学效应打下了基础.
Objective: To select suitable conditions for prokaryotic expression and purification of rhIL-17. Methods: rhIL-17 was expressed in E. coli host under heat induction. After compared among the expression amounts in different media under different heat induction time, the most suitable conditions was selected. The target protein was present in the form of inclusion body. The precipitate of inclusion was obtained and purified after 6M guanidine solublization or 2% SDS solublization. Results: Either protocol could yield rhIL-17 with high purity and stable activity. The SDS solublization mehthod gives rise to much more higher productivity than the guanidine solublization method. Conclusion: rhIL-17 were expression in E. coli system and purified to homogenicity by SDS solublization methods with high productivity.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
1999年第4期273-276,共4页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金重点项目(39730420)
国家杰出青年科学基金(39825123)资助
关键词
原核表达
纯化
鉴定
IL-17
γhIL-17
recombinant human interlukin 17
prokaryotic expression
renaturation
purification
characterization
