鼠高迁移率族蛋白1A盒基因的克隆和原核表达  

Cloning and expression of mouse HMGB1 a box gene

作　　者：姜南艳[1] 于文彬[2] 张惠中[1] 郝晓柯[2] 苏明权[2] 李斌[1] 李立文[3] 

机构地区：[1]第四军医大学唐都医院检验科,西安710038 [2]第四军医大学西京医院检验科 [3]第四军医大学西京医院全军骨科中心

出　　处：《陕西医学杂志》2007年第3期272-273,299,共3页Shaanxi Medical Journal

基　　金：西省科技攻关项目(2004K10-G4)

摘　　要：目的:克隆小鼠高迁移率族蛋白1A盒(HMGB1 A box)cDNA,并在大肠杆菌中表达GST-A盒融合蛋白。方法:从鼠肺提取总RNA,经RT-PCR获得HMGB1 A盒基因。将该基因插入pGEX-4T-2载体的BamHI和EcoRI位点之间并测序鉴定,转化BL21(DE3)后30℃IPTG诱导表达5h,进行SDS-PAGE分析。结果:DNA测序证明,获得了HMGB1 A盒基因,其序列与GenBank中报道序列基本一致。SDS-PAGE分析表明,HMGB1 A盒与GST融合蛋白获得成功表达,分子质量约36KD,表达量约占菌体总蛋白的15%,结论:通过RT-PCR成功克隆和表达了鼠HMGB1 A盒基因。Objective： To clone, express mouse High mobility group box chromosomal protein 1 （HMGB1） A box gene. Methods： Total RNA was extracted from the lung tissue of a new born mouse, and the HMGB1 A box gene was obtained by RT-PCR using specific primers. Then the ／ box gene was cloned into BamHI and EcoRI site of pGEX-4T-2 expression vector and confirmed by sequencing. After transforming E. coli BL21（DE3）, the recombinant bacteria was induced at 30℃ for 5h, the fusion protein GST-A box was analyzed by SDS-PAGE. Results： DNA sequencing result showed that HMGB1 A box gene was exactly consistent with the sequence reported in GenBank. SDS-PAGE analysis demonstrated that HMGB1 A-GST protein was expressed in E. coli BL21 （DE3）, and the molecular mass of it is 36 KD. The protein band amounted to 15 % of total bacteria protein. Conclusion： Mouse HMGB1 A box gene was successfully cloned and expressed.

关 键 词：HMGB1A盒/免疫学 大肠杆菌/免疫学 聚合酶链反应 序列分析 克隆 分子 基因表达 小鼠 

分 类 号：R563[医药卫生—呼吸系统]