Def-2,-3,-6 and-8, novel mouse genes differentially expressed in the haemopoietic system
Def-2,-3,-6 and-8, novel mouse genes differentially expressed in the haemopoietic system
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DOI：
10.1046/j.1365-2141.1999.01551.x
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发表时间：
1999-08-01
影响因子：
6.5
通讯作者：
Sablitzky, F
中科院分区：
文献类型：
--
作者：
Hotfilder, M;Baxendale, S;Sablitzky, F
为了鉴定髓系分化过程中发育调控的基因，使用携带β-半乳糖苷酶-新霉素的自失活逆转录病毒基因捕获载体，构建SA/lacZ/neo融合基因，并将其感染骨髓祖细胞（FDCP-混合物A4），G418抗性和β-半乳糖苷酶阳性细胞系（基因捕获整合[GTI]克隆）并诱导其在体外分化成巨噬细胞或粒细胞，通过分析成熟细胞类型的β-半乳糖苷酶活性，在单细胞水平上监测捕获位点的表达。测试的所有37个GTI克隆在粒细胞或粒细胞和巨噬细胞分化期间均显示下调。从8个克隆中分离与SA/lacZ/neo报告基因融合的内源编码区。分子生物学分析显示，其中一半代表新的小鼠基因（def-2，-3，-6和-8），我们证实了在初级造血组织中的差异表达。数据库检索显示def-2（与造血祖细胞相关）和def-8（在外周血白细胞中表达最强）没有显著相似性。Def-g在分化成髓系和红系谱系时被下调，被发现与最近描述的B细胞特异性开关重组酶SWAP-70密切相关但不相同。Def-3在分化成粒细胞时下调，但在祖细胞和巨噬细胞中表达，定义了一个新的RNA结合蛋白家族。
To identify developmentally regulated genes during myeloid differentiation, a self-inactivating retroviral gene-trap vector carrying a beta-galactnsidase-neomycin (SA/lacZ/neo) fusion gene was constructed and used to infect myeloid progenitor cells (FDCP-Mix A4), G418-resistant and beta-galactosidase positive cell lines (gene-trap integration [GTI] clones) were established and induced to differentiate in vitro into either macrophages or granulocytes, Expression of the trapped loci was monitored at a single-cell level by analysing the mature cell types for beta-galactosidase activity. All 37 GTI clones tested showed down-regulation either during granulocyte or both granulocytic and macrophage differentiation. The endogenous coding regions fused to the SA/lacZ/neo reporter gene were isolated from eight clones. Molecular analysis revealed that half of them represented novel mouse genes (def-2, -3, -6 and -8) which we confirmed to be differentially expressed in primary haemopoietic tissues. Database searches revealed no significant similarities for def-2 (associated with haemopoietic progenitors) and def-8 (expressed most strongly in peripheral leucocytes). Def-g, which is down-regulated upon the differentiation into myeloid as well as erythroid lineages, was found to be closely related but not identical with the recently described B-cell-specific switch recombinase SWAP-70. Def-3, which is downregulated upon differentiation into granulocytes but expressed in progenitor cells and macrophages, defines a novel family of RNA binding proteins.