3-1E/mChIL-15融合基因构建及在真核细胞中的表达  

Construction of eukaryotic expressing plasmid of E. acervulina 3-1E/ mChIL-15 fusion gene and its expression in eukaryotic cells

作　　者：马德星[1] 潘龙[1] 郎跃深[2] 杨静红[1] 李广兴[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]河北秦皇岛青龙职教中心,河北秦皇岛066500

出　　处：《东北农业大学学报》2010年第8期54-59,共6页Journal of Northeast Agricultural University

基　　金：国家自然科学基金(30901061);黑龙江省自然科学基金重点项目(ZNJ0702-01)

摘　　要：将鸡堆形艾美尔球虫（E.acervulina）子孢子和裂殖子表面抗原3-1E基因片段与鸡白细胞介素15成熟蛋白基因片段（mChIL15）通过四个柔性氨基酸SPGS连接,构建并鉴定真核表达质粒pcDNA3.1/3-1E-linker-mChIL-15。表达质粒纯化后应用磷酸钙法体外转染293T细胞,通过间接免疫荧光和免疫组织化学方法对重组质粒的体外瞬时表达进行检测。结果表明,成功构建了融合基因3-1E-linker-mChIL-15,转染后30h可检测到融合基因编码的融合蛋白在293T细胞中的瞬时表达。研究为鸡艾美尔球虫基因工程疫苗进一步研制及应用奠定了基础。The 3-1E antigen gene fragment located on the surface of E. acervulina sporozoites and schizonts was linked with mature chicken interleukin 15 （mChIL-15） protein gene by fourflexible amino acid SPGS to construct an expression plasmid pcDNA3.1/3-1E-mChIL-15. The plasmid was transfected into 293T cells in vitro with Ca3（PO4）2. The transient expression of the fusion protein was detected by indirect immunofluorescence assay and immunohistochemistry. The results showed that the fusion gene 3-1E-mChIL-15 was successfully constructed. The transient expression product of this fusion gene could be detected after transfection in 293T cell at 30 h. This research paves the way for the further study and application of Eimeria genetic engineering vaccine.

关 键 词：3-1E/mChIL-15 融合基因 真核表达质粒 瞬时表达 

分 类 号：Q78[生物学—分子生物学] Q503