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目的构建乙型肝炎病毒(HBV)颗粒性抗原转基因番茄植株。方法将乙型肝炎病毒preS/S基因插入到HBc的第73～94aa处(棘突尖部),再把整个复合基因克隆到植物表达载体pBIN438(含35s起动子)中,通过液氮冻融法转化根瘤农杆菌eha105,采用叶盘法转化番茄。结果得到一批转基因番茄植株,经PCR、PCR-Southernblot和Souhternblot证实HBc-HBs基因已整合到番茄基因组中;经Westernblot证实HBc-HBs能在番茄中有效表达。结论成功构建了乙肝颗粒性抗原基因的转基因番茄植株,为下一步进行该番茄疫苗株的效果评价奠定了基础。
Objective To construct hepatitis B virus (HBV) particle antigen transgenic tomato plants. Methods Hepatitis B virus preS / S gene was inserted into the 73 ~ 94 aa of the HBc (the tip of the spinous process), and then the entire complex gene was cloned into the plant expression vector pBIN438 (containing 35s promoter) Act to transform Agrobacterium tumefaciens eha105, using the leaf disc method tomato transformation. As a result, a batch of transgenic tomato plants were obtained. The results of PCR-Southern blot and Souhternblot showed that the HBc-HBs gene was integrated into the genome of tomato. Western blot showed that HBc-HBs could be efficiently expressed in tomato. Conclusion The transgenic tomato plants with hepatitis B particle antigen gene were successfully constructed, which laid the foundation for the next step to evaluate the effect of this tomato vaccine strain.