逆转录病毒介导重组CD23 cDNA 稳定整合的RVc23-8866细胞株的建立

建立稳定整合的细胞株，为研究外源基因定位整合机制及其对靶细胞生物特征影响，建立一个生物模型。方法构建重组病毒载体ＰＬＸＣＤ；用Ｌｉｐｏｆｅｃｔｉｎ将ＰＬＸＣＤ转民装细胞，用Ｇ４５１８筛选获得抗性细胞克隆，扩增抗性细胞收集病毒上清；测定病毒上清效价，感染ＲＰＭＩ８８６６细胞，用Ｇ４１８筛选获得抗性细胞；

Establishment of RVc23-8866 cell line transfected by retrovirus mediated recombinant CD23 cDNA

Objective:To establish a stable cell line and employ the cell line as a model to study target integration of exogenous cDNA and modulation of gene expression.Methods:Establish recombinant retrovirus vector PLXCD;use PLXCD to transfect PA317 cells by lipofectin,screen the transfected cells by G418,culture the cells to get supernatant containing viruses;measure the titer of the virus containing supernatant and transfect RPMI 8866 cells,screen the RVc23 8866 cells by G418;analyze the expression of CD23 on the membrane of RVc23 8866 by FACS;analyze the change of genome by In Situ Hybridization (FISH Method).Results:A stable cell line,RVc23 8866 was established.Conclusions:The results of FISH indicate that the recombinant PLXCD has integrated into the genome of RVc23 8866 cells;the results of FACS indicate that the expression of RVc23 8866 cells;the results of FACS indicate that the expression of membranal CD23 diminished and the integrated CD23 cDNA fragment has effected the expression of inherent CD23 gene.