应用反转录聚合酶链式反应检测鸡Th1样淋巴因子mRNA表达的研究

A Reverse Transcription_polymerase Chain Reaction(RT_PCR)Method was used to Analyze Chicken Thl_like lymphokines Gene

应用反转录聚合酶链式反应(RT_PCR) 和限制性酶切分析技术对鸡脾细胞体外受到有丝分裂原ConA刺激的情况下,其Th1 样淋巴因子mRNA的表达水平进行了研究。实验结果表明,ConA诱导培养3 小时的鸡脾细胞表达α_干扰素(ChIFN_α) 、γ_干扰素(ChIFN_γ)和白细胞介素_2(ChIL_2) 等鸡的Th1 样淋巴因子mRNA。未诱导但培养了3 小时的鸡脾细胞可表达ChIFN_α和ChIFN_γmRNA,而未检测到ChIL_2m RNA的表达。正常SPF鸡的脾细胞可表达ChIFN_αmRNA。本研究应用不同公司生产的反转录酶和Taq 多聚酶,取得了相同的结果。表明该方法具有很好的重复性,而且该法敏感而方便。

A reverse transcription_polymerase chain reaction(RT_PCR) and restriction enzyme digestion were used to analyze expression of genes coding for chicken Th1_like lymphokines in ConA_stimulated splenocytes. It was demonstrated that the IFN_α mRNA was expressed in the ConA_stimulated and unstimulated splenocytes after 3 hours in vitro cultivation.Expession of IFN_γ mRNA was detected in ConA_stimulated splenocytes and unstimulated splenocytes which cultured for 3 hours, but it was not detectable in the unstimulated splenocytes uncultured. Chicken IL_2 mRNA was only detected in ConA_stimulated splenocytes but not detected in other conditions. Results indicated that RT_PCR and restriction enzyme digestion were sensitive and convenient methods to monitor lymphokines mRNA expression in chicken at various conditions.