广西眼镜蛇神经生长因子的克隆及序列分析

CLONING AND SEQUENCING OF NERVE GROWTH FACTOR GENE FROM Naja naja atra

目的:扩增广西眼镜蛇神经生长因子(NGF)的全长基因并进行序列分析。方法:应用聚合酶链反应技术,以广西眼镜蛇cDNA为模板,扩增出编码NGF的全长基因,并克隆至pMD18-T载体中进行基因测序。结果:所得序列与Genbank提供的序列(M61180)对比显示,广西眼镜蛇prepro-NGF与Naja kaouthia(monocled cobra)、Bungarus multicinctus、mouse、人的NGF同源性分别为99%、91%、66%、68%,将不同氨基酸相同的化学性质计算在内,同源性更高。结论:成功地获得了广西眼镜蛇NGF的全长基因,且序列与已知序列高度一致。

Objective： To amplify and sequence nerve growth factor （NGF） gene from Naja naja atra. Methods： NGF was obtained by polymerase chain reaction by using Naja naja atra genomic DNA as a template and cloned into pMD18-T vector and sequenced. Result： The sequence of NGF was compared with those of the other snake venom（Naja kaouthia （monocled cobra） ,Bungarus multicinctus,mouse and human. The proteases of the sequence were aligned to maximize mutual homologies （99 %, 91 %,66 %, 68 % respectively）. Conclusion. NGF gene from Naja naja atra was successfully obtained and its sequence was highly consistent with the ones reported.