| 汪宗桂，左长清，刘振杰,刘新光.C11orf17蛋白的原核表达及功能预测.[J].中南医学科学杂志.,2014,42(6):541-544. |
| C11orf17蛋白的原核表达及功能预测 |
| Prokaryotic Expression and Function Prediction of C11orf17 Protein |
| 投稿时间：2013-11-02 |
| DOI： |
| 中文关键词: C11orf17 原核表达 生物信息学 |
| 英文关键词:C11orf17 prokaryotic expression bioinformatics |
| 基金项目:国家自然科学基金(30871440)，广东省自然科学基金重点项目(9252402301000002)，湛江市科技计划项目（2010C3106003），广东医学院科研基金面上项目（M2010005）. |
|
| 摘要点击次数: 996 |
| 全文下载次数: 1139 |
| 中文摘要: |
| 目的利用基因工程技术获取原核表达的C11orf17蛋白，结合生物信息学初步预测其功能。方法以K562细胞的cDNA为模板，PCR扩增C11orf17的基因序列；双酶切后将目的基因亚克隆至原核表达载体pET-32a上，将测序正确的重组表达质粒pET-32a-C11orf17转化到E.coli BL21，经诱导后SDS-PAGE及Western blot检测融合蛋白表达，并分析其可溶性。采用UGET软件预测C11orf17共表达基因，蛋白质互作数据库挖掘C11orf17基因已知互作蛋白，在线软件Gather富集共表达基因生物功能。结果成功构建重组表达质粒pET-32a-C11orf17，此重组体经诱导后能在E.coli BL21高效表达C11orf17的融合蛋白，UGET分析C11orf17前300共表达探针中，包含267个已知基因，共表达基因主要富集细胞交流、信号传导、细胞周期基因功能本体。蛋白互作数据库表明，C11orf17与PRKACA蛋白互作，后者为细胞周期调控重要分子。结论在原核细胞中成功表达出C11orf17蛋白，生物信息预测其可能是细胞周期调控重要新分子，为后续研究提供了方向和线索。 |
| 英文摘要: |
| ObjectiveTo obtain C11orf17 by prokaryotic expression system and predict its function by bioinformatics.MethodsBreast cancer cell cDNA were synthesized as a template and the gene encoding C11orf17 was amplified by PCR,and PCR product was subcloned into expression vector pET-32a.The constructed recombinant plasmid pET-32a-C11orf17 was transformed to E.coli BL21 for expression under induction of IPTG.The expressed product was identified by SDS-PAGE and Western blot.The co-expressed gene of C11orf17 was predicted by UGET software and its interaction protein was excavated through protein interaction database.The enriching biological function of co-expressed genes was predicted by online software Gather.ResultsThe recombinant plasmid was constructed successfully and fusion protein was expressed in E.coli BL21 efficiently after induced by IPTG.The gene funcutional ontology such as cell communication,signal transduction and cell cycle were enriched among 267 co-expressed genes.The protein interaction database indicated that C11orf17 could interact with PRKACA,which was one important molucule for cell cycle regulation.ConclusionProtein C11orf17 was successfully expressed and it may be a novel cell cycle regulated protein. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |