鸡白细胞介素2真核表达质粒的构建、表达及对AIV疫苗免疫增强作用研究

Construction and expression of the eukaryotic expression vector pCI-ChIL-2-EGFP and enhancement of AIV vaccine

将鸡白细胞介素2(ChIL-2)基因亚克隆入pCI载体中,pEGFP-N1载体中的EGFP基因酶切后连入质粒pCI-ChIL-2中,构建真核表达质粒pCI-ChIL-2-EGFP,该载体表达ChIL-2-EGFP融合蛋白。重组质粒在脂质体作用下转染Vero细胞、鸡胚成纤维细胞(CEF)和外周血淋巴细胞(PBLC),荧光显微镜观察到融合蛋白在体外能稳定表达。HI试验显示,pCI-ChIL-2-EGFP与AIV疫苗共注射可明显提高AIV疫苗的免疫原性。

Chicken interleukin-2 （ChlL-2） and enhanced green fluorescence protein （ EGFP ） gene were cloned into eu- karyotic expression pCI vector, resulting in pCI-ChIL-2-EGFP, which encode a fusion protein （ChIL-2-EGFP）. Vero cell, chicken embryo fibroblasts （CEF）, and periphery blood lymphocytes （PBLC） were transfected with pCI-ChIL-2-EGFP and Lipofectaine 2000. The expression of ChIL-2-EGFP fusion protein was found in the three cell strains under fluorescent microscope. The result of HI assay indicated that the efficacy of AIV vaccine could be modulated by co-administration of pCI- ChlL-2- EGFP plasmid.