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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。
Objective: To obtain recombinant human interleukin-15 (rhIL 1 5) highly expressed strains. Methods: Total RNA was extracted from human peripheral blood mononuclear cells activated by bacterial lipopolysaccharide + γ interferon. The gene fragment encoding human IL-15 cDNA was amplified by RT-PCR. The recombinant plasmid pBV220 was used to amplify the recombinant plasmid. Construction of IL15 gene engineering bacteria. Results: The results of nucleic acid sequencing were consistent with the expected results. SDS-PAGE showed that the molecular weight of IL-15 was about 15kD and the expression level was 28% of the total bacterial proteins. The CTL12 cells showed that the crude extracts were 1:1 0 0 refolding titer can reach 106IU / ml. Conclusion: The constructed genetically engineered bacteria are highly expressed strains of IL-15.