以pET-SUMO质粒为载体的凋亡素组合肽基因工程菌的构建  

Construction of genetically engineered bacteria for apoptin combination peptide with pET-SUMO plasmid as carrier

作　　者：闫佳敏 韩雪剑 张慧姝 谭培珠 魏惠妍 谢辉 王懿 李梦楠 张丽秋 YAN Jia-min;HAN Xue-jian;ZHANG Hui-shu;TAN Pei-zhu;WEI Hui-yan;XIE Hui;WANG Yi;LI Meng-nan;ZHANG Li-qiu(Biotechnology Experimental Teaching Center,School of Basic Medical Sciences,Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学基础医学院生物技术实验教学中心,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2022年第1期88-90,共3页Journal of Harbin Medical University

摘　　要：目的构建以pET-SUMO为载体的凋亡素组合肽基因工程菌,建立简便易行、能够放大的重组凋亡素组合肽纯化方法和工序。方法以pET-SUMO质粒为表达载体,构建凋亡素组合肽基因工程菌,通过镍柱纯化、SDS PAGE、小肽电泳检测重组凋亡素组合肽表达水平,选取适合转化、生产需求的基因工程菌株。结果测序证实凋亡素组合肽与pET-SUMO成功重组,第78~209碱基为凋亡素组合肽的编码序列,200 mmol/L咪唑洗脱融合蛋白表达水平高,收率高。结论确定pET-SUMO凋亡素组合肽基因工程菌为凋亡素组合肽转化开发的备用菌株。Objective To construct a genetically engineered strain of apoptin combination peptide with pET-SUMO as a carrier,and establish a simple,easy and scalable purification method and procedure for recombinant apoptin combination peptide.Methods pET-SUMO plas-mide was used to construct a genetically engineered apoptin combination peptide,and the expression of apoptin combination peptide was detected by the nickel column purification,SDS-PAGE,small molecules electrophoresis to select genetically engineered strains suitable for transformation and production.Results Sequencing showed that apoptin combination peptide was successfully recombined with pET-SUMO,Base 78th〜209th were the coding sequence of apoptin combination peptide.The 200 mmol/L imidazole eluted fusion protein had high expression level and high yield.Conclusion pET-SUMO apoptin combination peptide genetic engineering strain is a standby strain developed for the transformation of apoptin combination peptide.

关 键 词：凋亡素 凋亡素组合肽 基因工程 亲和层析 

分 类 号：R392.11[医药卫生—免疫学]