2种重组真核蛋氨酸酶原核体系表达及活性比较  

Expression and Activities of Two Prokaryotic Expression Vectors of Eukaryotic L-methionine γ-lyase Genes

作　　者：田腊梅[1] 任小丽[1] 胡振良[1] 曾红静 田长富[1] 

机构地区：[1]昆明医科大学生物医学工程研究中心基因与蛋白质工程研究室,云南昆明650500

出　　处：《昆明医科大学学报》2012年第5期11-14,共5页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(307660287);云南省科技厅重点基金资助项目(2009CC022)

摘　　要：目的构建2种真核L-蛋氨酸γ-裂解酶(MGL1 MGL2)的高效原核表达载体,诱导表达及纯化,比较活性差异.方法通过分子克隆技术构建2种重组表达质粒pGEX-4T-1-MGL1,pGEX-4T-1-MGL2;重组质粒转化感受态大肠杆菌Dh5α,诱导表达纯化蛋氨酸酶.结果构建了高效表达载体pGEX-4T-1-MGL1,pGEX-4T-1-MGL2;pGEX-4T-1-MGL1表达的酶纯度为82.5%,活性为0.505 2 IU/mg,pGEX-4T-1-MGL2表达的酶纯度为81.4%,活性为0.305 2 IU/mg.结论 pGEX-4T-1-MGL1,pGEX-4T-1-MGL2都能表达蛋氨酸酶;pGEX-4T-1-MGL1表达的蛋氨酸酶纯度比较高,酶活性比较高.Objective To construct two highly efficient prokaryotic expression vectors of eukaryotic L-methionine γ-lyase genes(MGL1 MGL2),induce their expression,purify,and compare their activities.Methods Two types of recombinant plasmids pGEX-4T-1-MGL1 and pGEX-4T-1-MGL2 were established by molecular cloning technique.Then the recombinant plasmids were transformed into E.coli Dh5ct to induce expression.Results Two types of highly efficient recombinant expression plasmids pGEX-4T-1-MGL1 and pGEX-4T-1-MGL2 were constructed successfully.Purity of metase of pGEX-4T-1-MGL1 was 82.5%,activity was 0.5052 IU/mg.Purity of metase of pGEX-4T-1-MGL2 was 81.4%,activity was 0.3052 IU/mg.Conclusion PGEX-4 T-1-MGL1 and pGEX-4 T-1-MGL2 can express L-methionine γ-lyase,and L-methionine γ-lyase of PGEX-4 T-1-MGL1 is purer and more active.

关 键 词：肿瘤 蛋氨酸酶 基因治疗 

分 类 号：Q782[生物学—分子生物学]