人白细胞介素10的克隆及分泌表达  

The cloning and secreting expression of human interleukin-10 in E.coli

作　　者：杨红[1] 罗少洪[1] 彭立胜[2] 钟肖芬[2] 

机构地区：[1]广东药学院生化教研室,广东广州510224 [2]中山大学生命科学学院,广东广州510275

出　　处：《广东药学院学报》2003年第1期48-50,共3页Academic Journal of Guangdong College of Pharmacy

基　　金：广州市科技局重大科技攻关项目

摘　　要：目的 :研究人白细胞介素 10 (hIL -10 )的高效分泌表达。方法 :根据Genbank报道的hIL -10基因序列 ,人工合成hIL -10基因 ,并将某些稀有密码子替换成E .Coli偏爱密码子 ,克隆至 pUC18质粒中 ,测序结果与预期一致。将hIL -10基因克隆至PET2 2b ,构建分泌表达克隆PET2 2b -hIL -10 ,经IPTG诱导在大肠杆菌BL2 1(DE3 )中表达hIL -10天然蛋白。结果 :SDS -PGAE电泳及凝胶密度扫描分析显示 ,重组蛋白分子量约为 18KD和 2 1KD(含信号肽 ) ,不同温度条件下重组蛋白的表达状态和表达量不同。结论 :18℃重组蛋白hIL -10的表达量为 7.6%。Objective: To study the secreting expression of human interleukin-10(hIL-10). Methods: According to the reported human interleukin-10 sequence,hIL-10 gene was synthesized and replaced some rare codes with hobby codes. The hIL-10 gene was inserted into PUC18 plasmid,identified by DNA sequencing. The expression plasmid referred to as PET22b-hIL-10 was constructed by inserting the cloned hIL-10 genes into secreting expression vector PET22b. The hIL-10 was overexpressed induced by IPTG in E.coli BL21. Results: Detected by densitometry analysis,the MW of recombinant protein was about 18KD and 21KD(with signal peptide),and its expression level varied at different temperatures. Conclusion: The expression level of the recombinant hIL-10 is about 7.6% at 18 ℃.

关 键 词：人白细胞介素10 克隆 分泌表达 

分 类 号：R392[医药卫生—免疫学]