摘要
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 establish recombinant tomato for HBV particle antigen.Method Hepatitis B virus preS/S gene was inserted into the 73～94aa of the HBc(tine of the spinous tuber). Then the whole combined gene was cloned into plant expression vector pBIN438 which is under the control of the CaMV 35s promotor. Tomato was transformed by co cultivating leaf discs with Agrobacterum strains harboring pBIN438 HBc HBs.Result The regenerated kanamycin resistant transforms were analyzed by PCR ,PCR Southern blot,Southern blot and Western blot. Conclusion The results indicated that HBc HBs was integrated into the genomic DNA of the tomato, and was efficiently expressed in the tomato.
出处
《热带医学杂志》
CAS
2004年第3期234-236,278,共4页
Journal of Tropical Medicine
基金
国家自然科学基金项目(No.30170532)
广州市重点攻关项目(No.2001-Z-035-01-2)。
