中国兽医科学2025，Vol.55Issue(5)：640-644,5.DOI:10.16656/j.issn.1673-4696.2025.0081
链球菌溶血素O真核表达载体构建及重组蛋白的表达纯化
Construction of eukaryotic expression vector for streptolysin O and purification of recombinant expression protein
摘要
Abstract
To obtain an antigen protein close to its natural form,the streptolysin O(SLO)gene was synthesized based on the sequence published in GenBank and cloned into the eukaryotic vector pPICZαA,construct pPICZαA-SLO recombinant plasmid.After identification by enzyme digestion and sequencing,the recombinant plasmid was transformed into Escherichia coli DH5α competent cells.The correct expression strain of Pichia pastoris was constructed by PCR identification and gene sequencing,and the recombinant protein was identified after purification using Ni-NTA affinity chromatography technology.The results showed that the Pichia sp GS115(pPICZαA-SLO)expression strain of Pichia pastoris was successfully constructed.The target protein with a relative molecular weight of 65 kDa was identified and detected by SDS-PAGE and Western-blot,indicating that SLO was successfully expressed and purified in Pichia sp,this study lays the foundation for the industrial production of SLO protein.关键词
链球菌溶血素O/真核表达/蛋白表达/纯化Key words
streptolysin O/eukaryotic expression/protein expression/purification分类
农业科技引用本文复制引用
向祝,徐莹,刘意,赵格,高玉斌,刘俊辉,张喜悦,王君玮..链球菌溶血素O真核表达载体构建及重组蛋白的表达纯化[J].中国兽医科学,2025,55(5):640-644,5.基金项目
国家重点研发计划项目(2022YFC2303900,2022YFD1301003) （2022YFC2303900,2022YFD1301003）
中国动物卫生与流行病学中心创新基金项目(DW2021001-13) （DW2021001-13）
