Objective To construct and identify retroviral-mediated short hairpin RNA ( shRNA ) expression vectors of ERβ419, and explore ERβ419 unknown biological function in beagles in future.Methods To screen out the most effective gene silencing sequence of beagle ERβ419 mRNA using qRT-PCR and Western Blot assays, imitate beagle estrogen target cells.Results qRT-PCR results showed, ERβ419-shRNA1 ( P <0.01 ) and ERβ419-shRNA3 ( P <0.01)differed significantly, Western Blot result as same as qRT-PCR,ERβ419-shRNA3 is the best choice.Conclusion Beagles ERβ419-shRNA3 retrain most effectively target gene repression. It is applied to explore ERβ419 unknown biological function in beagles reproductive system, and to prevent and treat beagles reproductive function diseases.%目的：构建最有效的慢病毒介导RNAi干扰序列，未来将应用于比格犬ERβ419的未知生物学功能的探索。方法模拟比格犬ERβ419靶细胞筛选针对目的基因mRNA设计的最佳干扰序列实验，通过qRT-PCR和Western Blot技术筛选最佳沉默表达载体序列。结果 qRT-PCR 结果显示， ERβ419-shRNA1（ P ＜0.01）和ERβ419-shRNA3（P ＜0.01）的差异表达极显著，Western Blot结果与qRT-PCR结果一致，ERβ419-shRNA3的干扰效果最佳。结论 ERβ419-shRNA3最有效地抑制了目的基因的表达，未来将应用于探索比格犬ERβ419未知生物学功能对比格犬生殖系统影响的研究，并进而预防和治疗比格犬繁殖机能障碍性疾病。
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