摘要
沉默信息调节因子2(Sir2)在染色质沉默、基因调控、代谢调节和调节细胞寿命等一系列细胞生物活动过程中起重要作用,近年来被作为抗病原微生物药物靶标成为了研究的热点。本研究应用生物信息学和比较基因组学技术注释获得了柔嫩艾美耳球虫(E.tenella)Sir2的基因序列,以注释的基因序列设计引物,以E.tenella第2代裂殖子cDNA文库为模板,利用PCR扩增获得了EtSir2ORF基因序列;并构建pET43a-EtSir2重组表达载体,进行原核表达。结果显示,EtSir2ORF全长为909bp,编码一段全长为302个氨基酸的多肽片段,该融合蛋白分子质量约为99ku,与预期分子质量一致。
Silent information regulator 2（Sir2）proteins play an important role in a wide array of functions like chromatin silencing,gene regulation,metabolic regulation,longevity and a series of cell biological activities.Pharmaceuticals that target with Sir2may be useful in resisting pathogenic microbes and thus become research focus in recent years.Sir2gene sequence of Eimeria tenella was obtained by gene annotation depended on comparative genomics and bioinformatics technology in present study.A potential EtSir2ORF was amplified and subcloned into the pET43a-EtSir2expression vector for prokaryotic expression.Our data indicated that,the full length of EtSir2ORF was 909bp which encodes 302amino acids with a predicted molecular mass of 99ku,and in accordance with expect.In summary,the present study successfully expressed and purified EtSir2,which established foundation for anti-coccidian drug screening in terms of EtSir2target.
出处
《动物医学进展》
CSCD
北大核心
2013年第7期39-43,共5页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(31201902)
珠江科技新星项目(2012J2200059)
广东省农业攻关项目(2012A020100001)
