重组人源Ⅲ型胶原蛋白在大肠杆菌中的表达与纯化工艺  

Expression and Purification Process of Recombinant Human Type Ⅲ Collagen in Escherichia Coli

作　　者：徐兰举[1] 柴雪晴 刘语菲 赵强[2] 刘鑫 李华 王淑芳[2] 杨卓[1] Xu Lanju;Chai Xueqing;Liu Yufei;Zhao Qiang;Liu Xin;Li Hua;Wang Shufang;Yang Zhuo(School of Medicine,Nankai University,Tianjin 300071,China;College of Life Sciences,Nankai University,Tianjin 300071,China;Hebei NACOL Biotechnology Co.,Ltd.,Shijiazhuang O50000,China)

机构地区：[1]南开大学医学院,天津300071 [2]南开大学生命科学学院,天津300071 [3]河北纳科生物科技有限公司,河北石家庄050000

出　　处：《南开大学学报（自然科学版）》2024年第3期40-45,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市研究生科研创新项目(2020YJSB118);天津市科技支撑重点项目(20YFZCSY01020)。

摘　　要：通过基因工程法优化人胶原蛋白的密码子序列,以人Ⅲ型胶原蛋白α1链为模板,(Gly-X-Y)为最小研究单位,构建重组质粒pET30a(+)-3CH,利用Escherichia coli发酵和蛋白纯化技术制备重组人Ⅲ型胶原蛋白(Recombinant human type Ⅲ collagen,RHCⅢ).研究结果证实,RHCⅢ的表达量为3.5 g/L,利用亲和/离子交换层析工艺纯化后的RHCⅢ纯度为95.44%,RHCⅢ蛋白溶液在4℃下可以保存12个月,稳定性良好,无细胞毒性,为重组人胶原蛋白产业化奠定了良好基础.The codon sequence of human collagen was optimized by genetic engineering method.Using theαl chain of human collagen Ⅲ as the template and(Gly-X-Y)as the minimum research unit,recombinant plasmid PET3a(+)-3CH,Escherichia coli fermentation and protein purification technology were used to prepare recombinant human type Ⅲ collagen(RHC Ⅲ).The results confirmed that the expression level of RHC Ⅲ was about 3.5 g/L,and the purity of RHC Ⅲ was about 95.44% after purification by affinity/ion exchange chromatography.The RHC Ⅲ protein solution could be stored at 4℃ for 12 months,with good stability and no cytotoxicity.The results laid a good foundation for the industrialization of recombinant human collagen.

关 键 词：基因工程 重组人Ⅲ型胶原蛋白 大肠杆菌 蛋白纯化 

分 类 号：Q812[生物学—生物工程]