博尔纳病病毒磷蛋白细胞模型的构建与鉴定

Construction and identification Borna disease virus phosphoprotein cell model

目的建立博尔纳病病毒(Borna disease virus,BDV)磷蛋白的细胞模型,并对所建模型进行鉴定。方法重新扩增和鉴定已有的BDV磷蛋白(GFP-P24)质粒,使用转染试剂将该质粒转入PC12细胞,并用荧光定量PCR和ELISA的方法对所构建的细胞模型进行鉴定。结果重新扩增的质粒PCR鉴定阳性,其浓度符合转染的需要,测序后未发现核苷酸的突变;转染PC12细胞的效率高,荧光定量PCR和ELISA检测均为阳性。结论成功构建了BDV磷蛋白的细胞模型,为研究BDV感染过程中磷蛋白所起的作用奠定了基础。

Objective To construct and identify the Borna disease virus （BDV） Phosphoprotein （P24） cell mod- el. Method BDV Phosphoprotein plasmids after amplification was verified by FQ-PCR, Agarose gel eleetrophoresis and Sequencing. BDV P24 was expressed in PC12 by transfection. The cell model was identified by FQ-PCR and ELISA. Result The BDV P24 plasmids meet the needs of transfeetion in concentration and no mutation was found by Sequencing. The rate of transfection in PC12 was high and both PCR and ELISA test ＇were positive. Conclusion P24 model was well constructed in PC12 which laid the foundation for further study of BDV.