摘要
高迁移率族蛋白是细胞"警报素"的一员,具有多样的生物学效应。本研究利用生物信息学技术从柔嫩艾美耳球虫转录组中鉴定到Et HMGB1基因序列,根据获得的基因序列设计引物,以柔嫩艾美耳球虫未孢子化卵囊总RNA为模板,通过RT-PCR技术获得Et HMGB1基因,并构建p ET28a-Et HMGB1重组表达载体,进行原核表达。结果显示,Et HMGB1基因全长为432 bp,编码1段全长为143个氨基酸的多肽,该融合蛋白分子质量约为16 000,与预期分子质量一致。Et HMGB1基因的成功克隆和表达为该蛋白的功能研究奠定了基础。
High mobility group protein is one of cell alarmins member, which have variety of biolog- ical effects. A putative HMGB1 gene sequence of E. tenella was obtained from E. tenella transcrip- tome by gene annotation depended on bioinformatics technology in present study. A potential EtH- MGB10RF was amplified by RT-PCR and subcloned into the pET28a-EtHMGB1 expression vec- tor for prokaryotic expression. The result showed that the full length of EtHMGB10RF was 432 bp,which encodes 143 amino acids with a predicted molecular mass of 16 000,and conformed to the expect. In conclusion, our study successfully expressed EtHMGB1 in E. coli which laid a foundation for its function study.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2016年第4期613-616,共4页
Chinese Journal of Veterinary Science
基金
安徽省教育厅重大资助项目(KJ2014ZD09)
安徽省教育厅重点资助项目(KJ2015A189)
安徽科技学院校级重点资助项目(ZRC2014406)
安徽科技学院重点学科资助项目(AKXK20101-2)