通过对小鼠补体受体Ⅱ型基因（CR2）进行定点突变，然后将野生型和突变型小鼠CR2/1(MCR2/1)及人CR2(hCR2)用电穿孔方法导入小鼠SP2/0进行表达，采用免疫组织化学等方法鉴定阳性细胞系。用EB病毒对转染阳性细胞进行感染，EBER-1杂交结果显示，仅转染hCR2和突变型MCR2（mtMCR2）的SP2/0细胞感染了EB病毒，前者感染EB病毒的阳性率较后者高很多。这为进一步研究EB病毒进入细胞的机制及建立EB病毒相关的鼻咽癌动物模型奠定了良好的基础。%Site-directed mutagenesis method was used to introduce two desired mutations, which were confirmed by DNA sequencing, into mouse complement receptor Type II gene(MCR2). Then the constructed eukaryotic expression vectors containing wild type mouse CR2/1(wtMCR2/1), mutant type mouse CR2/1 (mtMCR2/1) and human CR2 (hCR2) cDNA were transferred into mouse SP2/0 cells by electroporation. After two-week screening by G418, the stably transfected clones were obtained. Several ways including PCR, RT-PCR, and immunohistochemistry were utilized to screen those clones with interesting genes integrated and expressed. Then Epstein-Barr virus(EBV) was used to infect these transfected cells and EBER-1 (EBV encoded RNAs) hybridization results showed that only hCR2 and mtMCR2 transfected SP2/0 cells could be infected by EBV, but positive rate of the former was much higher than the latter. This study sets groundwork for elucidating the mechanism by which EBV enters the cells and for establishing the animal model of EBV-related nasopharyngeal carcinoma (NPC).
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