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克隆小鼠白细胞介素 18(IL 18)编码区的cDNA ,并实现在真核细胞中的表达。方法 :用小鼠白细胞介素 18(mIL 18)的cDNA核苷酸序列 ,设计、合成基因序列特异性引物 ,应用逆转录多聚酶链反应RT PCR ,以植物血凝 (PHA)和细菌脂多糖 (LPS)刺激的小鼠非粘附性脾细胞的mRNA为模板 ,扩增获得全长mIL 18,转染小鼠成纤维细胞系SVT2 ,并进行IL 18生物学活性的检测。结果 :经测序证实获得的小鼠IL 18的cDNA序列与文献报道的mIL 18的cDNA序列完全一致。构建的小鼠IL 18的重组表达载体在转染小鼠成纤维细胞SVT2后 ,获得具有生物学活性mIL 18的分泌表达。结论 :克隆了小鼠IL 18的cDNA ,并实现了在真核细胞中的表达。
Mouse cDNA for the interleukin 18 (IL 18) coding region was cloned and expressed in eukaryotic cells. Methods: The cDNA sequence of mouse IL - 18 was designed and synthesized. Reverse transcriptase - polymerase chain reaction (RT - PCR) and reverse transcription - polymerase chain reaction (RT - PCR) LPS) stimulated mouse non-adherent spleen cells as a template, amplified to obtain full-length mIL 18, transfected mouse fibroblast cell line SVT2, and the detection of IL 18 biological activity. Results: The cDNA sequence of mouse IL 18 obtained by sequencing was exactly the same as the cDNA sequence of mIL 18 reported in the literature. Recombinant Expression Vector for Constructed Mouse IL 18 [0184] Secreted expression of biologically active mIL 18 was obtained following transfection of mouse fibroblast SVT2. Conclusion: cDNA of mouse IL 18 was cloned and expressed in eukaryotic cells.