CAJ | 학술논문

克隆人胰岛素原基因，通过密码子优化和融合蛋白化学水解位点和纯化标签优化，构建人胰岛素原高效原核表达系统，为重组人胰岛素原的高效表达纯化制备奠定基础。根据大肠杆菌密码子偏好性优化设计人胰岛素原基因，通过PCR技术获得重组基因，构建原核表达载体pET-32a-proinsulin，转化大肠杆菌表达株E．coli BL21，筛选人胰岛素原高效表达菌株，建立诱导表达和纯化融合蛋白技术方法。克隆获得优化的人胰岛素原重组基因，构建原核高效表达载体，获得优化的诱导表达方法和纯化的人胰岛素原。该研究为建立胰岛素高效制备的优化条件奠定基础。
극륭인이도소원기인，통과밀마자우화화융합단백화학수해위점화순화표첨우화，구건인이도소원고효원핵표체계통，위중조인이도소원적고효표체순화제비전정기출。근거대장간균밀마자편호성우화설계인이도소원기인，통과PCR기술획득중조기인，구건원핵표체재체pET-32a-proinsulin，전화대장간균표체주E．coli BL21，사선인이도소원고효표체균주，건립유도표체화순화융합단백기술방법。극륭획득우화적인이도소원중조기인，구건원핵고효표체재체，획득우화적유도표체방법화순화적인이도소원。해연구위건립이도소고효제비적우화조건전정기출。
To optimize human proinsulin codon , fusion protein chemical hydrolysis sites and purification tags and construct human proinsulin prokaryotic expression system for high efficient expression and purifica -tion.Methods:Designing proinsulin gene according to the Escherichia coli password preference .Getting the pro-insulin gene by PCR technology and construct the cloning prokaryotic expression vector pET -32a-proinsulin. The pET-32a-proinsulin vector was imported into E.coli BL21.Screening human proinsulin high expression strain,establishing the method for proinsulin induced expression and fusion protein purification .The optimiza-tion proinsulin gene was obtained , a high effective prokaryotic expression vector was constructed , the better method for proinsulin expression and purified human proinsulin were obtained .The study provides the founda-tion for increasing insulin output .