乙型肝炎病毒表面抗原基因在墨西哥酸浆中的表达及其免疫原性的初步研究

以墨西哥酸浆子叶作为外植体，与农杆菌EHA105（含质粒p130HBs)共培养3天，通过延迟筛选的方法用潮霉素加压直接诱导成芽，抗性芽经诱导生根获得转化植株。经PCR、Southern点杂交等分子检测，证实目的基因已整合到墨西哥酸浆基因组中。ELISA检测证实了在墨西哥酸浆中表达的HBsAg具有较好的活性。初步比较了目的基因在转化植株、不同器官的表达水平差异。取注射注免1次但抗体水平明显下降的Babl/c小鼠，口服饲喂加强免疫，有明显的特异性抗体回升反应。表明转基因墨西哥酸浆疫苗作为口服加强免疫具有重要的理论和实用价值。

The Oral Vaccination of HBV by Cherry Tomatillo transformed with HBsAg

Cotyledons of cherry tomatillo were co-cultured with Agrobacterium tumefaciens strain EHA105(harbouring p1301HBs) for three days.Through hygromycin delay-selection,transformed cherry tomatillos were obtained.The presence and integration of foreign DNA in transgenic cherry tomatillo was confirmed by hygromycin resistance selection,Gus detection,polymerase chain reaction(PCR) and dot blot analysis.The immunoactivity of recombinant HBsAg(rHBsAg) was identified by ELISA.The expression levels of rHBsAg in different organs of transgenic cherry tomatillos were compared.Immunogenicity of rHBsAg derived from transgenic cherry tomatillo tissues was confirmed by oral feed of transformed tissues to Balb/c mice primed with commercial HBV vaccines.Specific serum antibody was boosted in this case.The feasibility of transgenic cherry tomatillo as oral vaccine is proved,and this study will provide some theoretic and experimental directions for the production of large-scale,low-cost oral HBV vaccines using transgenic cherry tomatillo.