兔转基因单细胞克隆株的分离培养及其染色体倍性分析

为检测原代二倍体细胞转基因后单细胞克隆的增殖能力及其染色体倍性稳定性，用脂质体介导的转染方法将质粒DNA pEGFP-C1（带有报告基因GFP和Neo^r）导入体外培养的兔胎儿成纤维细胞中，经G418药物筛选后，分离出73个GFP阳性细胞克隆，最后存活13个（18％），对其中9个克隆的染色体倍性进行分析，结果只有2个（22％）克隆的染色体倍性正常率在75％以上，分别为80％和75％，其余7个克隆的染色体倍性正常率均在70％以上。这表明，当使用转基因单细胞克隆株作为供核细胞产生克隆动物时，单细胞克隆的增殖代数和染色体倍性的稳定性需要进一步研究提高。

Cell Cloning from Transfected Rabbit Primary Cell Lines and Analysis of Their Ploidy

In order to examine the viability and ploidy of cell clones derived from transfected primary cell lines, the plasmid pEGFP-C1 (containing both reporter gene gfp and neo) were delivered into rabbit fetal fibroblast cells cultured in vitro using the method mediated by lipofectamine. After selection under G418, 73 clones were picked out, of which only 13(18%)survived the sequent culture. When chromosome counting of cells from 9 clones were performed, it was found that only 2(22%)clones have a normal chromosome number, with 80% and 75% diploid cells in their population respectively. In remaining 7 clones the percentage of the cells which have normal chromosome number was less than 70%. These results indicates that when transgenic cell clones were used as donor cells to produce cloned animal by nuclear transfer, further study is needed to improve the viability and ploidy stability of cell clones.