柔嫩艾美耳球虫HMGB1基因的克隆与原核表达

Cloning of Eimeria tenella high mobility group box1 gene and its expression in E.coli

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.