Zhang X T, YU D M, WU H L
College of Life Science, Reking University, Beijing.
Yi Chuan Xue Bao. 1998 Dec;25(6):491-8.
A MT-II gene targeting vector--pMT-II6.7 with 6.7kb sequences homolog to mMT-II and its flanking region has been transfected into Mespu22 with electroporation. We have got 26 positive clones from 104 G418 and Ganc clones with PCR method. From the karyotype analysis, we found two clones with 84% and 88% normal karyotype. They are clone 5-2 and 8-4. Using Southern analysis, we confirm that the two clones are homologous recombinants. These cell lines still have the pluropotential ability to differentiate both in vitro and in vivo. Now we have got a chimera mouse with cells from clone 8-4.
一个与小鼠金属硫蛋白-II(mMT-II)及其侧翼区域具有6.7kb同源序列的MT-II基因靶向载体——pMT-II6.7,已通过电穿孔法转染到Mespu22细胞中。我们用PCR方法从104个G418和Ganc抗性克隆中获得了26个阳性克隆。通过核型分析,我们发现两个克隆(克隆5-2和8-4)的核型正常率分别为84%和88%。通过Southern分析,我们证实这两个克隆是同源重组体。这些细胞系在体内外仍具有多能分化能力。现在我们已经获得了一只嵌合体小鼠,其细胞来源于克隆8-4。