腺病毒载体介导的GFP基因在鸭胚成纤维细胞中的表达及RNA干扰

目的：建立在鸭胚成纤维细胞（DEF）中进行RNA干扰（RNAi）的技术平台，为鸭基因组功能的研究提供新的技术手段。方法：以绿色荧光蛋白（GFP）基因为报告基因，脂质体转染化学合成的GFP特异小干扰RNA（GFP-siRNA），用流式细胞仪测定GFP-siRNA对重组腺病毒（Adv-GFP）介导的GFP基因在DEF中表达的干扰效果。结果：200MOI（感染复数）Adv-GFP介导的GFP基因在DEF中表达效率最高，为31．20％±3．1l％，对DEF的活力无明显影响；GFP-siRNA能有效干扰GFP基因在DEF中的表达，相对抑制率为98．56％。结论：在DEF中进行RNAi是可行的，Adv-GFP是介导外源基因在DEF中表达较为理想的载体；首次建立了在DEF中进行RNAi的技术平台，为鸭基因组的功能等研究提供了新的技术手段。

Expression of GFP Gene Mediated by Adenovirus Vector and RNA Interference in Duck Embryo Fibroblast

Objective： To establish a methodological platform for RNA interference（RNAi） studies in duck embryo fibroblast（DEF）, so as to provide a novel way for the studies of genome functions of ducks. Methods： Green fluorescent protein （GFP） gene was used as report gene and DEF was transfected by synthesized anti-GFP small interfering RNA（siRNA） （GFP-siRNA） using the method of cationic liposome reagent. Interference effect of GFP-siRNA to GFP gene expression that was mediated by adenovirus（Adv-GFP） in DEF was evaluated by flow cytometer method. Results 200 multiplicity of infection（MOI） Adv-GFP could get highest expression rates of GFP gene（31.20%±3.11%）, and wouldn＇t influence cell activity of DEF. GFP-siRNA could significantly interfere with the expression of GFP gene in the DEF, and the relative inhibitory rate was 98.56%. Conclusion： The results showed that it is feasible to carry out RNAi in DEF and Adv-GFP is an ideal vector for exogenous gene expression in DEF. The present study established a methodological platform for RNAi studies in DEF for the first time, and provided a novel way for the studies of genome functions of ducks.