摘要
目的合成重组大鼠高迁移率族蛋白B1 A结构域(HMGB1-A)蛋白。方法利用基因工程技术,克隆并重组大鼠高迁移率族蛋白B1 A盒基因;利用原核表达技术,表达并纯化重组大鼠HMGB1-A盒蛋白,并利用Western blot法进行验证。结果琼脂糖凝胶电泳分析显示HMGB1-A盒基因大小约为250 bp;重组克隆p UC-A经双酶切鉴定,产物大小约为3000 bp、250 bp;重组克隆p GEX-A PCR产物大小约为250 bp。原核表达产物Mr大小约为36 000,与预期一致。结论成功表达并纯化HMGB1-A盒蛋白。
Objective To obtain the recombinant A-domain of high mobility group box 1（ HMGB1-A） protein.Methods Using genetic engineering techniques, we cloned and recombined rat HMGB1-A gene. Using prokaryotic expression technique,we expressed and purified the recombinant rat HMGB1-A protein,which was verified by Western blotting. Results Agarose gel electrophoresis analysis showed that HMGB1-A cassette gene size was about 250 bp; double digestion for identifying the recombinant clone p UC-A showed that the product size was about 3000 bp,250 bp; PCR for identifying the recombinant clone p GEX-A showed that the product size was about 250 bp. SDS-PAGE revealed that the product size was about 36 000,which was consistent with the expectation. Conclusion The recombinant rat HMGB1-A was successfully expressed and purified.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2017年第9期1201-1205,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
陕西省社会发展攻关计划项目(2016SF-405)
