Generation of a conditional allele of the B-myb gene
Generation of a conditional allele of the B-myb gene
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DOI：
10.1002/gene.20170
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发表时间：
2005-12-01
期刊：
影响因子：
1.5
通讯作者：
Frampton, J
中科院分区：
文献类型：
--
作者：
García, P;Berlanga, O;Frampton, J
B-Myb是一种重要的转录因子，在多种细胞类型中参与控制细胞周期和调节组织特异性基因的表达。这两个等位基因的缺失会导致E4.5-6.5的早期胚胎死亡。为了研究B-Myb在胚胎发育后期和特定成体组织中的功能，我们产生了一个B-myb等位基因(B-mybF)。用转基因GATA-Cre小鼠品系进行饲养，证实了Cre介导的体内缺失。在Pgk-neo(R)盒存在于内含子3(B-MYB(Loxneo))的小花等位基因的产生过程中产生的一个中间等位基因，被推测为一种弱低晶型，与B-MYB(-/-)胚胎相比，纯合子的胚胎死亡较晚。为了证明B-MYB失活的有效性和可能的后果，我们对培养的MEF进行了条件缺失，并观察到与异常核DNA复制相关的生长减少。
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.