鸭MCHR2的基因克隆及蛋白表达鉴定

Gene cloning and protein expression identification of duck MCHR2

目的筛查能够稳定表达黑色素浓集激素受体2(MCHR2)基因的动物,为构建动物模型提供实验依据。方法利用逆转录-多聚酶链反应(RT-PCR)方法,从新鲜的鸭、Wista鼠、SD鼠、C3H10鼠、C57鼠、狗、羊、猪等大脑组织中提取总RNA,以此扩增MCHR2基因的全长cDNA编码区序列;利用Western blot检测鸭、犬和猪的大脑海马区及皮质区MCHR2蛋白的表达情况。结果通过RT-PCR获得鸭、犬、猪脑组织中的目的片段;通过Western blot验证了MCHR2蛋白在鸭脑组织中具有高表达。结论将鸭作为构建动物模型的对象,用于MCHR2基因的相关功能研究具有可行性。

Aim To provide evidence for animal model construction by screening for animals with a stable expression of MCHR2. Methods The full-length MCHR2 eDNA fragment was amplified by RT-PCR from the total RNA of fresh brain tissue of duck, SD rat, C3H10 mice, C57 mice, Wista mice, dog, goat and pig. Western blot was used to detect the expression of MCHR2 protein in brain tissue of duck, as well as the cortex and hippoeampus of dog and pig. Results After obtainning the full-length MCHR2 eDNA fragment of the brain tissue of duck, dog and pig by RT-PCR, the high expression of MCHR2 protein in brain tissue of duck by Western blot was confirmed. Conclusion It is feasible to use duck as animal model for further research on MCHR2 function.