轮状病毒VP4-ST融合基因转化小球藻的筛选

Selection of chlorella transformed with rotavirus VP4-ST fusion gene

进行了椭圆小球藻对G418和卡那霉素的抗生素敏感性试验,同时采用电激转化法将构建好的含35S启动子的重组植物表达载体pBin-VP4-ST转化椭圆小球藻,并对转基因小球藻进行了PCR和PCR-Southern-blot检测。结果显示,30mg/L的G418为转化子的最佳抗生素筛选浓度。PCR和PCR-South-ern-blot检测结果显示,VP4-ST已成功插入到转基因小球藻基因组中,从而为用转基因小球藻作为生物反应器生产抗腹泻口服疫苗奠定了基础。

The sensitivity tests of Chlorella ellipsoidea against kanamycin and G418 were conducted, and the re-constructed vector pBin-VP4-ST was transformed into chlorella through electroporation. The transgenic chlorellas were identified by PCR and PCR-Southern-blot. The results showed that the optimal selection condition of transformed ehlorella was G418 30 mg/L, and the positive transformed ehlorella of the VP4-ST fusion gene was obtained. These results established a foundation for using transgenic chlorella as an efficient bioreactor system to produce an oral anti-diarrhea vaccine.