重组质粒PcDNA3．1-P30-P22-CTXA2／B在Hela细胞中表达的研究

目的观察重组质粒 pcDNA3.1-P30-P22-CTXA_2/B 在 Hela 细胞中的表达情况,并检测其表达的融合蛋白 P30-P22-CTXA_2/B 免疫学活性。方法脂质体介导重组真核表达质粒 pcDNA3.1-P30-P22-CTXA_2/B 转染 Hela 细胞,G418筛选稳定转染的细胞株,收集蛋白行 SDS-PAGE 和 Westem-Blot 分析,检测融合蛋白的免疫学活性。结果经重组质粒 pcDNA3.1-P30-P22-CTXA_2/B 稳定转染的 Hela 细胞表达了分子量约64KD 的融合蛋白,经 Western-Blot 检测证实该融合蛋白具有 P30免疫原性。结论在HeLa 细胞中成功表达了具有 P30免疫原性的融合蛋白 P30-P22-CTXA_2/B,为下一步动物实验奠定了基础。

Experiment study of recombinant plasmid pcDNA3.1 - P30 - P22 - CTXA2/B expressing in Hela cells

Objective To study the expression of recombinant plasmid pcDNA3.1-P30-P22-CTXA_2/B in Hela cells,and detect the immunological activity of fusion protein P30-P22-CTXA_2/B.Method Hela cells were transfected with recombinant plasmid pcDNA3.1-P30-P22-CTXA_2/B by liposomes.After screened with G418, the expressed products of P30-P22-CTXA_2/B in the transfected Hela cells were detected by running SDS-PAGE and assayed by Western-Blot.Results P30-P22-CTXA_2/B complex gene were highly expressed in stable trans- fectants and the fusion protein about 64KD with obvious P30 immunological activity.Conclusion Fusion protein with obvious immunological activity was successfully expressed in Hela cells.This would pave way for the next experiments on mice.