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从鸭外周血分离的单核细胞在体外经PHA (5 μg/ml)刺激 2 0h后 ,提取总RNA及mRNA并通过逆转录制备cDNA。采用最近克隆的鸡白细胞介素 2 (ChIL 2 )cDNA序列为探针对cDNA进行Southernblot杂交 ,以识别鸭白细胞介素 2 (DuIL 2 )特异的cDNA序列。从鸭外周血单核细胞中提取的基因组DNA经限制性内切酶BamHI、EcoRI、XbaI消化后 ,以上述探针进行杂交 ,分析DuIL 2基因组序列。结果表明 ,无论是鸭cDNA或基因组DNA ,经Southern杂交后 ,均得到特异性杂交条带 ,证实ChIL 2与DuIL 2具有较高序列同源性。DuIL 2mRNA表达时形成两种转录子 ,大小分别为 95 9bp、 780bp。同鸡γ 干扰素(ChIFN γ )探针产生的杂交信号比较 ,ChIL 2产生的杂交信号相对较弱 ,推测可能和DuIL 2mRNA在上述条件下拷贝数相对较少以及ChIL 2与DuIL 2cDNA序列同源性相对较低有关
Monocytes isolated from peripheral blood of ducklings were stimulated in vitro with PHA (5 μg / ml) for 20 h. Total RNA and mRNA were extracted and cDNA was prepared by reverse transcription. The recently cloned chicken interleukin - 2 (ChIL 2) cDNA sequence was used as a probe to Southern blot hybridization of cDNA to identify duck IL - 2 specific cDNA sequence. Genomic DNA extracted from duck peripheral blood mononuclear cells was digested with restriction enzymes BamHI, EcoRI, and XbaI, and hybridized with the above probe to analyze the DuIL 2 genome sequence. The results showed that the specific hybridization bands of both duck cDNA or genomic DNA were obtained after Southern hybridization, which confirmed that ChIL 2 had high sequence homology with DuIL 2. DuIL 2 mRNA forms two transcripts when expressed, with sizes of 95 9 bp and 780 bp, respectively. The hybridization signal generated by ChIL 2 is relatively weaker than the hybridization signal generated by the chicken IFNγ (ChIFN γ) probe, suggesting that the copy number of DuIL 2 mRNA may be relatively low under the above conditions and the homology of the ChIL 2 and DuIL 2 cDNA sequences Relatively low sexual related