INACTIVATING THE BETA-2-MICROGLOBULIN LOCUS IN MOUSE EMBRYONIC STEM-CELLS BY HOMOLOGOUS RECOMBINATION
INACTIVATING THE BETA-2-MICROGLOBULIN LOCUS IN MOUSE EMBRYONIC STEM-CELLS BY HOMOLOGOUS RECOMBINATION
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DOI：
10.1073/pnas.86.22.8932
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发表时间：
1989-11-01
影响因子：
11.1
通讯作者：
SMITHIES, O
中科院分区：
文献类型：
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作者：
KOLLER, BH;SMITHIES, O
通过基因打靶，我们已经灭活了小鼠胚胎干细胞系中的内源性β2-微球蛋白基因。将编码序列被新霉素抗性基因插入打乱的β2-微球蛋白基因的克隆片段用于转染胚胎干细胞。选择G418抗性菌落，然后用聚合酶链式反应筛选，以鉴定那些传入的DNA通过同源重组整合到胚胎干细胞基因组中的菌落。在筛选的234个G418抗性菌落中，鉴定出2个正确的靶标菌落。从这两个靶向胚胎细胞系中都获得了携带灭活β2-微球蛋白基因的嵌合小鼠。培育这种动物的后代将有助于研究β2-微球蛋白纯合缺失的影响。
We have inactivated, by gene targeting, the endogenous .beta.2-microglobulin gene in a mouse embryonic stem cell line. A cloned fragment of the .beta.2-microglobulin gene with the coding sequence disrupted by the insertion of the neomycin-resistance gene was used to transfect the embryonic stem cells. G418-resistant colonies were selected and then screened using the polymerase chain reaction to identify those in which the incoming DNA had integrated into the embryonic stem cell genome by homologous recombination. Of a total of 234 G418-resistant colonies screened, 2 correctly targeted colonies were identified. Chimeric mice carrying the inactivated .beta.2-microglobulin gene have been obtained from both of these targeted embryonic cell lines. Breeding of offspring from such animals will allow investigation of the effects of homozygous loss of .beta.2-microglobulin.