摘要
目的 :获得重组人白细胞介素 1 5 (rhIL 1 5 )高效表达菌株。方法 :经细菌脂多糖 +γ干扰素活化的人外周血单个核细胞提取细胞总RNA ,用RT PCR方法扩增出编码人IL 1 5cDNA的基因片段 ,采用 pBV2 2 0表达载体 ,经DNA重组技术构建IL 1 5基因工程菌。结果 :核酸序列测定与预期一致 ,所表达的IL 1 5经SDS PAGE证明分子量约 1 5kD ,表达量占菌体总蛋白的 2 8% ,经CTLl2 细胞检测 ,表达产物粗提物 1∶1 0 0复性后效价可达到1 0 6IU /ml。结论 :构建的基因工程菌为IL 1 5高效表达菌株。
Purpose:To obtain human IL 15 high level expression vector.Methods:Human IL 15 cDNA fragments (0.35 kb)were amplified from LPS+IFNγ stimulated human monocytes by RT PCR method and inserted into EcoR I/BamH I site of pBV220 expression vector which contains P RP L promoters and SD sequence.Results:The expression level of rhIL 15 was more than 28% of the total bacterial protein in SDS PAGE,molecular weight was 14.4 kD.The biological activity of refolded bacterial extract(1∶100)was 10 6 IU/ml tested by using CTLl 2 cell line.Conclusion:Recombinant human IL 15 high level expression vector has been constructed in E.coli .
出处
《中国生化药物杂志》
CAS
CSCD
2000年第4期163-165,共3页
Chinese Journal of Biochemical Pharmaceutics
基金
山东省自然科学基金资助!(项目编号 :Y95C0 4 2 6)
