目的 利用转基因中药表达人细胞因子基因 ,探讨培育基因修饰 (GM)中药的可行性。方法 将体外扩增分别获得的人 α-干扰素基因与 RANTES基因酶切回收后插入中间载体 ,用 Eco R 和 H ind 双酶切鉴定重组子。由大肠杆菌分离重组质粒并导入根癌农杆菌 Agrobacterium tumefaciens,通过加入卡那霉素 (Km)和利福平(Rif)筛选含重组双元载体的转化子 ,用叶盘共培养法转化苦瓜和夏枯草外植体。结果 经 RT-PCR检测到人 α-干扰素基因与 RANTES基因在中药转化愈伤组织中的瞬时表达。结论 首次报道了重组人 α-干扰素基因与RANTES基因在转基因苦瓜和夏枯草细胞中的表达 ,为探讨利用转基因中药抗病毒尤其是抗艾滋病开创了新途径。

Object To explore the feasibility of breeding genetic modified (GM) medicine by expressing human cytokine in transgenic Chinese materia medica Methods Human interferon α gene and RANTES gene available from the amplification in vitro were enzymatically excised, recoveried, and inserted into intermediate vectors The recombinants were identified by double enzyme digestion of EcoRⅠand HindⅢ The plasmids were extracted from Escherichia coli and introduced into A tumefaciens, and the transformants harboring binary vectors were screened by addition of antibiotics of kanamycin (Km) and rifampicin (Rif), and the explants of M charantia and P vulgaris were transformed by co cultivation of leaf disks with A tumefaciens strain Results RT PCR was applied to detect the transient expression of human interferon α gene and RANTES gene in transformed medicinal herbal calli Conclusion The expression of recombinant human interferon α gene and RANTES gene in transgenic M charantia and P vulgaris cells was firstly reported, which opens an alternative road to antivirus, especially anti AIDS virus, by using transgenic Chinese materia medica

国家自然科学基金(39770902;39870725);广东省自然科学基金(98064);国家中医药管理局科研基金(97Z082)