分泌型rhGM-CSF/IL-2融合蛋白表达载体的构建  

Construction of Expression Vector of Secretory rhGM-CSF/IL-2 Fusion Protein

作　　者：刘冬梅[1] 张卓远 王妍[1] 孟洪琪[2] 

机构地区：[1]长春生物制品研究所,长春130062 [2]吉林大学公共卫生学院,长春130021

出　　处：《中国生物制品学杂志》2002年第4期197-200,共4页Chinese Journal of Biologicals

摘　　要：目的 将GM-CSF基因与IL-2基因通过一中间接头G-G-G-S-G-G-S-G连接到一起,并在E.coli中分泌表达具有更高GM-CSF和IL-2生物活性的融合蛋白分子。方法 在引物设计中,通过选择密码子,在Linker的中前部设计了一个BamHI酶切位点,GM-CSF的下游引物和IL-2的上游引物均起始于该位点。然后通过PCR、酶切、连接等一系列分子克隆的方法,构建融合蛋白的克隆及表达载体。结果 经EcoRI/SaII双酶切鉴定,克隆载体pUC19GM-CSF/L/IL-2及表达载体pBVGMCSF/L/IL-2中均插入了正确的外源片段。结论 克隆及表达载体的成功构建,为进一步表达融合蛋白及活性研究打下了基础。Objective To link GM-CSF gene to IL-2 gene by a mediate sequence G-G-G-S-G-G-S-G and express fusion protein with high biological activity in E. coli. Methods A BamHI digestion site from which the downstream primer of GM-CSF and upstream primer of IL-2 start was introduced to the front or middle part of the Linker, then the vectors for the cloning and expression of fusion protein were constructed by PCR, enzyme digestion and linkage. Results Restriction analysis with EcoRI/Sal I proved that the correct foreign fragments were inserted to the cloning vector pUC19GM - CSF/L/IL - 2 and expression vector pBVG-MCSF/L/IL-2. Conclusion The successful constructions of cloning and expression vectors laid a foundation of expression of fusion protein and study on the biological activity of it.

关 键 词：分泌型rhGM-CSF/IL-2融合蛋白 表达载体 构建 分泌表达 

分 类 号：R392.11[医药卫生—免疫学] R394.8[医药卫生—基础医学]