miR-200b-3p和miR-200b-5p在马立克氏病抗性与易感SPF鸡法氏囊组织的差异表达分析

摘要

Objective To validate high throughput sequencing results by fluorescence quantitative PCR technique and analyze the expression of miR-200b-3p and miR-200b-5p in bursa,and explore possible molecular mechanism associated with Marek's disease (MD) resistance/susceptibility.Methods Through optimization,the ideal amplification conditions and reaction system of real-time fluorescence quantitative PCR were identified.Results miR-200b-3p and miR-200b-5p fluorescence quantitative PCR results and high throughput sequencing results were consistent in down-regulation,but not significant difference.miR-200b-3p and miR-200b-5p two homologous miRNA showed similar expression after infected by Marek's disease virus (MDV).Conclusion Although the difference is not consistent with the results of significant analysis,miR-200b-3p and miR-200b-5p may be associated with resistance / susceptibility of MD.%目的 在前期高通量测序分析的基础上,进一步利用荧光定量PCR验证miR-200b-3p和miR-200b-5p在马立克氏病(MD)抗性(B21单倍型)与易感(B19单倍型)鸡法氏囊组织的差异表达情况.方法 通过经优化后确定的最佳实时荧光定量PCR反应体系和扩增条件,进行荧光定量PCR分析.结果 miR-200b-3p和miR-200b-5p荧光定量PCR结果与高通量测序结果上下调情况基本一致,但差异表达显著性分析结果却不完全相同.结论 尽管差异表达显著性分析结果不完全一致,但两种方法的结果均提示来源于同一前体的miR-200b-3p和miR-200b-5p可能与MD抗性/易感性相关.