【摘要】 为构建猪繁殖与呼吸综合征病毒受体CD163基因缺失细胞系和进一步明确CD163在PRRSV感染过程中的作用,本研究以猪基因组为模板,扩增猪繁殖与呼吸综合征病毒受体CD163基因同源左臂和同源右臂,并将扩增片段与pGEM-T载体连接后进行部分序列测定,同时与已知相应片段进行同源性分析,再将扩增的同源左右臂分别酶切后定向连接至pSSC-9载体中,成功构建了含Neo和Tk基因的双标记打靶载体pSSC-Larm-Sarm。更多还原

【Abstract】 In order to construct cell lines without CD163gene,agene of porcine reproductive and respiratory syndrome virus receptor,and find out role of CD163in PRRSV infection,5’and 3’homologous arms of CD163gene of porcine reproductive and respiratory syndrome virus receptor were amplified and cloned into pGEM-T vector.Results of sequencing showed that 5’and 3’homologous arms of CD163gene were 4.5kb and 1.1kb,respectively.Then,the homologous arms of CD163gene were subcloned into pSSC-9to construct a targeting vector pSSC-Larm-Sarm for CD163gene knock-out.The vector will be used to construct fibroblast cells with CD163gene knockout.更多还原