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目的克隆与小鼠红细胞终末分化相关因子的基因，并对其表达特性进行分析。方法用致贫血Ｆｒｉｅｎｄ病毒（ＦＶＡ）诱导ＢＡＬＢ／ｃ小鼠脾脏产生大量ＦＶＡ原红细胞，后者经分离纯化并在促红细胞生成素作用下进行体外培养。采用减除杂交与ＰＣＲ相结合的方法，用未经培养的ＦＶＡ诱导的原红细胞（０ｈＦＶＡ原红细胞）的ｃＤＮＡ对培养３６ｈ的ＦＶＡ诱导的成红细胞（３６ｈＦＶＡ成红细胞）的ｃＤＮＡ进行多次减除杂交和ＰＣＲ扩增，构建上述３６ｈｃＤＮＡ的质粒减除文库并进行差异筛选。对阳性克隆进行核苷酸序列测定及Ｎｏｒｔｈｅｒｎ印迹分析。结果获得１个４７２ｂｐ的ｃＤＮＡ片段，其中含有１个３０９ｂｐ的开放阅读框架，编码１０２个氨基酸。经ＧｅｎＢａｎｋ进行同源比较，未发现同源序列，已被ＧｅｎＢａｎｋ接受，（编号ＡＡ１１４３６９）。Ｎｏｒｔｈｅｒｎ印迹分析表明，该ｃＤＮＡ在培养３６ｈ的ＦＶＡ中、晚幼红细胞中呈差异表达，其ｍＲＮＡ全长约为５００～６００ｂｐ左右。将其暂命名为小鼠红细胞分化相关因子（ＭＥＤＲＦ）基因。结论ＭＥＤＲＦ基因在红细胞分化的中、晚幼阶段特异表达，表明ＭＥＤＲＦ是一个新的与红细胞终末分化相关的因子。
Objective To clone the genes related to mouse erythrocyte terminal differentiation and to analyze its expression characteristics. Methods FVA-induced erythrocytes were produced from the spleen of BALB / c mice by leukemia Friend virus (FVA), which was isolated and purified and cultured in vitro with erythropoietin. Using subtractive hybridization and PCR, cDNAs of FVA-induced erythrocytes (36hFVA erythrocytes) cultured for 36 hours were subtracted and hybridized several times with cDNA of non-cultured FVA-induced erythroblasts (0 hFVA erythroblasts) And PCR amplification to construct the 36h cDNA plasmid subtraction library and differential screening. Positive clones were subjected to nucleotide sequencing and Northern blot analysis. The results obtained a 472bp cDNA fragment, which contains a 309bp open reading frame encoding 102 amino acids. Homologous comparison by GenBank showed no homologous sequences and was accepted by GenBank (Accession No. AA114369). Northern blot analysis showed that the cDNA was differentially expressed in late-cultured erythrocytes in FVA cultured for 36 hours, and the total length of the cDNA was about 500-600 bp. This is temporarily called the mouse erythroid differentiation-related factor (MEDRF) gene. Conclusion The MEDRF gene is specifically expressed in the middle and late stages of erythroid differentiation, indicating that MEDRF is a new factor related to the terminal differentiation of erythrocytes.