A组轮状病毒结构蛋白基因在马铃薯细胞中的初步表达

目的 在马铃薯细胞中表达轮状病毒结构蛋白。方法采用RT-PCR扩增截短VP4蛋白基因，克隆于植物表达载体paBI221，再双酶切pBI221，将带有CaMV35S启动子、外源基因及终止子的片段转入pSB11，通过三亲杂交方法，制备农杆菌转化载体pSB111-VP4，并对pSB111-VP4作点杂交检测，筛选鉴定重组子，对马铃薯组织细胞进行转化。结果转化细胞经Western印迹检测其免疫活性，具有良好的抗原性。结论为研制轮状病毒重组疫苗、亚单组疫苗、转基因植物口服疫苗做了前期准备。

Expression of rotavirus structural protein gene in Potato

Objective Rotaviruses are the major pathogens that cause life threatening diarrhea in young children. The aim of this study is to express rotavirus structural protein in potato cells. Methods The partial gene of rotavirus VP 4 protein was amplified by RT-PCR, and cloned into pBI221.pBI221/VP 4 was digested with double enzymes and the little fragment containing promoter CaMV35S, foreign gene and terminator was transferred into pSB11. The recombination vector pSB111-VP 4 was constructed and selected by dot hybridization, and was transferred into potato cells. Results The immunocompetence of transferred cells was detected by western blot. The result showed that transferred cells had better antigenicity. Conclusion This study may provide a foundation for further research in the preparation of rotavirus transgene plant vaccine or oral rotavirus vaccine.