结肠特异性基因RELMβ的克隆及其真核表达载体构建

图1 小鼠各脏器组织中RELMβ的表达. A: Western blot; B: Northern blot. 1: 脑; 2: 心; 3: 肺; 4: 肝; 5: 脾; 6: 肾; 7: 胸腺; 8: 结肠; 9: 小肠; 10: 胃; 11: 肌肉; 12: 脂肪.

图2 小鼠结肠组织中RT-PCR扩增出RELMβ基因. M: 100 bp DNA marker; 1-8: PCR退火温度分别为65, 64.5, 63.3, 61.4, 58.9, 57.1, 55.8, 55℃.

图3 RELMβ基因核酸序列分析.

图4 真核表达载体pcDNA3. 1-RELMβ的鉴定. A: 酶切; M: 1 kb DNA marker; 1: pcDNA3.1/Zeo(+)/EcoRⅠ; 2-9: 重组子1-8/EcoRⅠ; B: PCR; M: 100 bp DNA marker; 1, 2, 4, 7: 反向插入的重组子; 3, 5, 6, 8: 正向插入的重组子.

图5 pcDNA3. 1-RELMβ在小鼠细胞中的表达. A: RT-PCR; B: Western blot; M: 100 bp DNA marker; 1: NIH 3T3对照组; 2: CT26对照组; 3: NIH 3T3 pcDNA3.1转染组; 4: CT26 pcDNA3.1转染组; 5: NIH 3T3 pcDNA3.1-RELMβ转染组; 6: CT26 pcDNA3.1-RELMβ转染组.