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目的 对表达轮状病毒VP7基因的 1株重组腺病毒rvAdG1VP7(G)的生物学特性进行鉴定 ,为进一步免疫学研究打下基础。方法 用rvAdG1VP7(G)感染 2 93细胞 ,并分别应用电子显微镜技术以及Southernblot、PCR、RT PCR和Westernblot等分子生物学方法了解rvAdG1VP7(G)的形态、VP7基因整合、遗传稳定性、VP7基因在细胞内转录及表达等有关生物学特性。结果 电子显微镜观察表明 :rvAdG1VP7(G)具有典型的腺病毒形态 ;Southernblot、PCR、RT PCR和Westernblot等分子生物学方法分析显示 :rvAdG1VP7(G)中确有特异性的VP7基因稳定整合 ,有较好的遗传稳定性 ,感染 2 93细胞后能有效转录 ,可表达轮状病毒主要结构蛋白VP7。结论 rvAdG1VP7(G)具有良好的生物学性质 ,为进行体内实验研究提供了必要条件。
Objective To identify the biological characteristics of a recombinant adenovirus rvAdG1VP7 (G) expressing the rotavirus VP7 gene and lay the foundation for further immunological research. Methods 293 cells were infected with rvAdG1VP7 (G), and the morphology of rvAdG1VP7 (G), the integration of VP7 gene and the genetic stability were analyzed by electron microscopy, Southern blot, PCR, RT PCR and Western blot respectively. Intracellular transcription and expression of the relevant biological characteristics. Results The results of electron microscopy showed that rvAdG1VP7 (G) had typical adenovirus morphology. Southern blot, PCR, RT-PCR and Western blot analysis showed that there was indeed a stable and integrated VP7 gene in rvAdG1VP7 (G) Good genetic stability, after infection of 293 cells can be effectively transcribed, the expression of rotavirus major structural protein VP7. Conclusion rvAdG1VP7 (G) has good biological properties, which provides the necessary conditions for in vivo experimental studies.