双基因导入人脐血CD34^+细胞共表达的调控

目的 观察携带双基因的逆转录病毒载体转导人脐血CD3 4+细胞后 ,猴病毒 4 0 (SV4 0 )早启动子和内部核糖体进入位点 (IRES)对载体上、下游双基因共表达的影响。方法 分别构建含SV4 0早启动子和IRES的双基因逆转录病毒载体pLESN和pLEIN ,两者均携带增强型绿色荧光蛋白和新霉素抗性neo基因。重组载体经包装以其病毒上清感染经磁性细胞分离仪富集的人脐血CD3 4+细胞 ,而后进行集落形成细胞测试。结果 流式细胞仪检测表明 10 %的脐血CD3 4+细胞表达绿色荧光蛋白。集落形成细胞测试发现pLESN转导的脐血CD3 4+细胞体外培养形成的G4 18抗性集落中 ,4 1 1%的集落有绿色荧光 ,而pLEIN组全部为绿色荧光集落。结论 与SV4 0早启动子相比 ,含有IRES序列的重组双基因逆转录病毒载体转导人脐血CD3 4+细胞时 ,能保证上、下游双基因的协同表达。

Incorporation of Resistance Gene into Tomato using Binary Vector System

Exoplants of tomato(Lycopersicon esculentum) leaf were transformed with Ti plasmids using binary vector system.After screening.with selection culture, kanamycin-resistant seedling were obtained from callus. Molecular hybridization proved the integration of Km gene into plant cell genome via A.tumefaciens. Higher activity of Nos-NPTase was demonstrated in the transformed plant,thus confirming the successful expression of the resistance gene in recipient cells.