摘要
应用DEAE-纤维素柱层析及Sephadex G-50柱层析纯化了天麻(Gastrodia elata Bl.)抗真菌蛋白(GAFP),并制备了多克隆抗体(GAFP-PcAbs)。通过蛋白质印迹杂交(Western blot)及点印迹杂交(Dotblot)证明GAFP-PcAbs为GAFP特异性抗体。进一步采用免疫荧光显微观察,对越冬前后新鲜次生天麻块茎作了GAFP的组织免疫荧光化学定位。结果表明GAFP定位于天麻次生块茎表皮和皮层,其中以表皮含量较高。
Gastrodia elata B1. antifungal protein (GAFP) was purfied by DEAE-Cellulase column chromatography and Sephadex G-50 column chromatography, and the polyclonal antibody against GAFP (GAFP-PcAbs) was also produced. Using Western blot and Dot blot it was demonstrated that GAFP-PcAbs is specific for GAFP. For further study, immunofluorescence localization of GAFP in secondry corms of G. elata has been conducted with fluorescence microscopy. The result showed that GAFP was localized at epidermal and cortical tissues in secondry corm of G. elata, and the content of GAFP was higher in epidermis.
关键词
天麻
抗真菌蛋白
免疫荧光
Gastrodia elata
Antifungal protein
Serological antigen
Western blot
Dot blot Immunofluorescence
