García Paloma, Berlanga Oscar, Watson Roger, Frampton Jon
Institute for Biomedical Research, Birmingham University Medical School, Edgbaston, United Kingdom.
Genesis. 2005 Dec;43(4):189-95. doi: 10.1002/gene.20170.
B-Myb is an essential transcription factor involved in control of the cell cycle and the regulation of tissue-specific gene expression in a wide range of cell types. Loss of both alleles results in early embryonic lethality at E4.5-6.5. To address the function of B-Myb in later stages of embryogenesis and in specific adult tissues, a floxed B-myb allele (B-mybF) was generated. Cre-mediated deletion in vivo was demonstrated by breeding with a transgenic GATA-Cre mouse line. An intermediate allele produced in the creation of the floxed allele, in which the PGK-neo(R) cassette is present in intron 3 (B-myb(loxneo)), was deduced to be a weak hypomorph based on the later embryonic death of homozygotes compared to B-myb(-/-) embryos. To demonstrate the efficiency and possible consequences of B-myb inactivation, we performed conditional deletion in cultured MEFs and observed decreased growth that correlated with aberrant nuclear DNA replication.
B-Myb是一种重要的转录因子,参与多种细胞类型的细胞周期调控和组织特异性基因表达的调节。两个等位基因的缺失会导致在E4.5-6.5时出现早期胚胎致死。为了研究B-Myb在胚胎发育后期和特定成年组织中的功能,构建了一个floxed B-myb等位基因(B-mybF)。通过与转基因GATA-Cre小鼠品系杂交,证明了体内Cre介导的缺失。在构建floxed等位基因过程中产生的一个中间等位基因,其中PGK-neo(R)盒存在于内含子3中(B-myb(loxneo)),根据纯合子与B-myb(-/-)胚胎相比后期胚胎死亡的情况,推断其为弱亚效等位基因。为了证明B-myb失活的效率和可能的后果,我们在培养的MEF中进行了条件性缺失,并观察到生长下降,这与异常的核DNA复制相关。