猪瘟病毒E2基因的哺乳动物细胞表达

利用PCR技术扩增出猪瘟病毒主要结构蛋白E2基因的全序列，将其克隆到真核表达载体PEGPF-C1中，获得重组质粒PEGPF-E2，经PCR，酶切鉴定和序列分析证明目的基因的大小、插入位置和读码框完全正确。利用脂质体将阳性质粒PEGFP-E2转染入猪肾来源的细胞PK-15中，G418筛选出阳性的细胞克隆，通过PCR证明E2基因存在于筛选出的阳性细胞克隆中。利用荧光倒置显微观察阳性细胞发现有融合蛋白的表达，利用夹心ELISA猪瘟病毒抗原检测试剂盒检测证明表达的融合蛋白为猪瘟病毒E2基因编码蛋白。

Expression E2 gene of Classical Swine Fever in Mammalian Cell

The major structure protein E2 gene of Classical swine fever was amplified by polymerase chain reaction (PCR).After the amplified fragments were cloned into the eukaryotic expression vector PEGPF-C1,the recombinant plasmids PEGPF-E2 were obtained. The insert position, the size and the reading frame were right by PCR, restriction digestion and the sequence analysis. The positive plasmids was transducted by liposome into PK-15 cell which origin from pig kidney. PCR indicated that E2 gene was in positive cell clone which selection by G418.Fusion protein expression observated by inverted fluorescence microscope. Classical Swine Fever Virus Antigen Test Kit detection indicated that express fusion protein is E2 gene code protein.