优化密码子提高草原兔尾鼠ZP3融合蛋白的原核表达  

Enhanced prokaryotic expression of a recombinant Lagurus lagurus zona pellucida subunit 3 with codon optimization

作　　者：李轶杰[1] 张富春[1] 张钰[1] 王焱冰[1] 

机构地区：[1]新疆大学生命科学与技术学院分子生物学重点实验室新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《细胞与分子免疫学杂志》2005年第6期700-703,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(No.30360062)

摘　　要：目的:探讨提高草原兔尾鼠(Lagurus lagurus)卵透明带3(LZP3)基因在原核细胞中表达的水平。方法:利用重叠PCR技术,定点突变LZP3基因上3个稀有的密码子簇,将LZP3基因中7个稀有的密码子更换成大肠杆菌(E.coli)最常用的相应密码子。将获得的LZP3突变基因(LZP3m)插入pGEX4T-1中,构建重组表达载体。以重组体转化E.coliBL21(DE3)菌株进行表达。结果:LZP3m基因的表达量比野生型LZP3基因明显提高。结论:通过密码子优化,能显著提高LZP3基因在原核细胞中的表达水平。AIM： To improve prokaryotic expression level of Lagurus lagurus zona pellucida 3 （ LZP3 ） gene. METHODS： The rare codons of LZP3 gene fragment were turned into the most frequently used codons in E. coil by overlap PCR. LZP3 gene and mutated LZP3 （LZP3m）gene were cloned into expression vector pGEX4T-1 respectively, and transformed into E. coil BE21 （DE3）. RESULTS. The expression level of LZP3m was notably higher than that of LZP3. CONCLUSION： The expression of LZP3 gene in E. coil could be successfully improved by using codon optimization.

关 键 词：草原兔尾鼠 卵透明带3 重叠PCR 密码子优化 原核表达 

分 类 号：Q786[生物学—分子生物学]