Co-expression of mic-5 gene of eimeria stiedai and rabbit il-15 in eucaryotic cell line | 斯氏艾美耳球虫MIC-5与兔IL-15基因在真核细胞中的共表达

Chinese. 目的构建能在真核细胞中共表达斯氏艾美耳球虫微线蛋白5（MIC-5）和兔IL-15重组载体，为进一步研究抗斯氏艾美耳球虫的免疫调节性DNA疫苗奠定基础。方法将斯氏艾美耳球虫MIC-5和兔IL-15基因，克隆到真核双表达载体pBudce4.1中，应用脂质体介导法将构建的重组质粒pB-MIC-5-IL-15转染BHK-21细胞。在转染后，用RT-PCR法和间接免疫荧光试验（IFA）分别检测MIC-5和IL-15基因在BHK-21细胞中的转录和表达情况。结果在重组质粒pB-MIC-5-IL-15转染BHK-21细胞24 h后即可检测到MIC-5、IL-15基因进行了转录；在转染36、48、72h后，MIC-5和IL-15基因可同时在BHK-21细胞中表达。结论成功构建了含有斯氏艾美耳球虫MIC-5和兔IL-15基因的重组载体，实现了两种基因在真核细胞中的共表达，为抗斯氏艾美耳球虫的免疫调节性DNA疫苗研制奠定了基础。

English. Objective The aim of this study is to construct a recombinant co-expression vector expressing Eimeria stiedai microneme protein 5 (MIC-5) and rabbit IL-15 in eukaryotic cells, which may lay a foundation for further study immune regulatory DNA vaccine against infection of Eimeria stiedai. Method E. stiedai MIC-5 gene and rabbit's IL-15 gene were respectively cloned into plasmid pBudce4.1 to construct recombinant vector pB-MIC-5-IL-15 for co-expression, and then the recombinant plasmid pB-MIC-5-IL-15 was transfected into BHK-21 cell line for expression. The RT-PCR and indirect fluorescent immunoassay were used to detect the transcription and expression of interesting genes in BHK-21 cell line, respectively. Result The results showed that E. stiedai MIC-5 gene and rabbit's IL-15 gene were successfully transcripted. And interesting genes of E.stiedai MIC-5 and rabbit's IL-15 were successfully expressed at 36, 48 and 72 hour post-transfection, respectively. Conclusion The recombinant co-expression vector was successfully constructed, which can express E. stiedai MIC-5 and rabbit IL-15 gene in eukaryotic cells.