Whitelaw C B, Springbett A J, Webster J, Clark J
AFRC-Institute of Animal Physiology and Genetics Research, Edinburgh Research Station, Midlothian, Scotland, UK.
Transgenic Res. 1993 Jan;2(1):29-32. doi: 10.1007/BF01977678.
Most transgenic mice are generated by the direct microinjection of DNA fragments into the pronuclei of fertilized eggs. It has been generally assumed that the majority of integration events occur prior to the first round of chromosomal DNA replication (Palmiter and Brinster, 1986). In this study we have determined by comparison of PCR, Southern blot and transmission frequencies that at least 62% of integration events generate a mosaic (somatic and/or germline) G0 transgenic mouse. Furthermore, the statistical probability of transgene-containing cells segregating to the various early embryo lineages implies that this is probably an underestimate of the true mosaic frequency. Thus, the majority of DNA injected into fertilized mouse eggs intergates after the first round of chromosomal DNA replication, therefore most G0 transgenic mice are derived from a mosaic embryo.
大多数转基因小鼠是通过将DNA片段直接显微注射到受精卵的原核中产生的。人们普遍认为,大多数整合事件发生在第一轮染色体DNA复制之前(帕尔米特和布林斯特,1986年)。在本研究中,我们通过比较PCR、Southern印迹和传递频率确定,至少62%的整合事件产生了嵌合(体细胞和/或生殖系)G0转基因小鼠。此外,含转基因细胞分离到各种早期胚胎谱系的统计概率表明,这可能低估了真正的嵌合频率。因此,注入受精卵的大多数DNA在第一轮染色体DNA复制后整合,因此大多数G0转基因小鼠来自嵌合胚胎。