人白介素17受体样分子的克隆及表达

目的　构建带有 6×myc的人白介素 17受体样分子的重组表达质粒 ,以便检测hIL 17RLM L基因在真核细胞中的特异性表达。方法　设计带有酶切位点 (EcoRⅠ和XhoⅠ )的特异性引物 ,用PCR方法扩增hIL 17RLM L片段回收后插入带有 6×myc标签的 pcDNA3.0真核表达载体 ,转染COS7细胞后作Westernblot检测其表达。结果　成功地构建了带有 6×myc标签的 pcDNA3.0 6×myc /hIL 17RLM L重组质粒 ,Westernblot检测到该质粒可在真核细胞中表达。结论　用分子生物学的方法成功地构建了真核表达质粒pcDNA3.0 6×myc /hIL 17RLM L ,使该基因的特异性检测成为可能 ,为进一步研究hIL 17RLM L的生物学功能奠定了基础。

The cloning and expression of human interleukin-17 receptor-like molecule

Objective To generate an human interleukin-17 receptor-like molecule (IL-17RLM) recombinant plasmid with 6×myc tag and detect its specific expression in eukaryotic cells. Methods Design two specific primers(including the enzyme sites of EcoRⅠand XhoⅠ), reextract hIL-17RLM-L DNA fragment after PCR and insert it into the 6×myc tagged pcDNA3.0 vector, then detect its expression by Western blot after transfecting COS7 cells. Results The 6×myc tagged recombinant plasmid pcDNA3.0- 6×myc /hIL-17RLM-L was generated successfully and its expression can be detected by Western blot in eukaryotic cells. Conclusion The eukaryotic expressing plasmid pcDNA3.0-6×myc /hIL-17RLM-L was generated successfully and its specific expression was realized, which may provide the basis for further research of its biological function.