免疫荧光法检测原核和真核细胞中表达的轮状病毒外壳蛋白VP4

目的：用免疫荧光法快速检测原核和真核细胞中表达的轮状病毒（RV）外壳蛋白VP4。方法：以抗VP4的抗体为一抗、FITC标记的羊抗豚鼠IgG为二抗，用免疫荧光方法检测在大肠杆菌BL21（DE3）中重组表达的同源RVVP4；检测SA11或Wa株RV感染MA104细胞后不同时间段病毒VP4的合成及其在感染细胞中的分布情况。结果：用免疫荧光法可直接检测到原核细胞中表达的外源蛋白，也可检测到病毒蛋白在真核细胞中的分布情况。结论：免疫荧光法可特异、方便、快速地检测RV VP4在原核和真核细胞中的表达；来源于RV TB—Chen株的VP4抗体可特异性识别同源病毒VP4，交叉识别SA11或Wa株的VP4。

Immunofluorescent Detection of Rotavirus Capsid Protein VP4 Expressed in Prokaryotic and Eukaryotic Cells

Objective： To detect the capsid protein VP4 of rotavirus（RV） expressed in prokaryotic and eukaryotic cells by immunofluorescent assay（IFA）. Methods： Using anti-recombinant RV VP4（rVP4） antibodies as detecting antibody and FITC-labled goat anti-guinea pig IgG as second antibody to detect rVP4 expressed in E.coli BL21 （DE3） cells and VP4 in MA104 cells infected by Wa- or SAil-stain of RV. Results： VP4 expressed in E.coli or in infected MA104 cells could be detected out by the IFA. Conclusion： IFA is a specific, rapid and convenient method for definition of RV VP4 and other virus proteins expression and their distribution in transformed or infected cells.