桃树李坏死环斑病毒的RT-PCR检测方法研究

Study on Detection of PNRSV in Peach by RT-PCR

以桃树叶片为材料提取总RNA,根据PNRSV外壳蛋白基因设计引物,进行RT和PCR扩增,获得良好结果,对目的片段经过克隆测序,与NCBI中报道的PNRSV外壳蛋白基因序列同源性最高达98%。在建立的RT-PCR技术基础上,对RT-PCR体系中的主要影响因子进行了优化研究。

Total RNA was extracted using peach leaves as material. Primers sequences to detect the PNRSV were designed according to the nucleotides sequences of the coat protein published in the Genbank database , NCBI. The targed amplication products were 449bp and the effect of RT-PCR were well. The specific fragment was cloned and its sequences were analyzed. The maximal homology of the two nucleotides sequences is 98% comparing the PCR products sequences with the reported se quences of PNRSV in database. Based on the established detection method of RT-PCR , the factor in RT-PCR system were optimized.