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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)
