Leprince D, Saule S, de Taisne C, Gegonne A, Begue A, Righi M, Stehelin D
EMBO J. 1983;2(7):1073-8. doi: 10.1002/j.1460-2075.1983.tb01548.x.
Chicken and human cell DNA contains sequences homologous to the avian myeloblastosis virus oncogene, v-myb. These cellular sequences, c-myb (human) and c-myb (chicken), were isolated from libraries of human or chicken cell DNA fragments, generated by partial digestion with the restriction enzymes AluI and HaeIII, and compared. The chicken c-myb locus isolated from two distinct overlapping recombinant phages, contained five contiguous EcoRI fragments of 5.4, 1.1, 2.1, 2.2 and 9 kbp, accounting for all the bands seen with a v-myb probe in a complete EcoRI digest of chicken cellular DNA. Likewise, the screening of the human library yielded a recombinant phage hybridizing with the v-myb specific probe, that contained five EcoRI fragments of 2.8, 2.6, 2.0, 1.2 and 5.0 kbp (the last ending with an artificial EcoRI site, due to the construction of the library) belonging to the c-myb (human) locus. Probes using the EcoRI chicken DNA cloned fragments revealed corresponding contiguous EcoRI fragments in the human clone. Subsequent analyses of cellular polyadenylated mRNA extracted from human and chicken cells allowed the identification of single RNA species of 3.8 and 4.0 kb, respectively, as the representative transcripts of the c-myb locus in the two species. Thus, c-myb appears as a single locus in man and chicken, conserved with a similar structure in the two distantly related species. Our preparation of a specific human c-myb probe with an increased sensitivity on DNA/RNA blots should facilitate analyses concerning this gene in human normal or tumour cells or tissues.
鸡和人类细胞的DNA含有与禽成髓细胞瘤病毒癌基因v-myb同源的序列。这些细胞序列,即c-myb(人类)和c-myb(鸡),是从用限制性内切酶AluI和HaeIII部分消化产生的人类或鸡细胞DNA片段文库中分离出来并进行比较的。从两个不同的重叠重组噬菌体中分离出的鸡c-myb基因座,包含5个连续的EcoRI片段,大小分别为5.4、1.1、2.1、2.2和9 kbp,这解释了在鸡细胞DNA的完整EcoRI消化物中用v-myb探针看到的所有条带。同样,对人类文库的筛选产生了一个与v-myb特异性探针杂交的重组噬菌体,它包含属于c-myb(人类)基因座的5个EcoRI片段,大小分别为2.8、2.6、2.0、1.2和5.0 kbp(最后一个片段由于文库构建而以人工EcoRI位点结束)。使用克隆的鸡DNA的EcoRI片段作为探针,在人类克隆中发现了相应的连续EcoRI片段。随后对从人类和鸡细胞中提取的细胞多聚腺苷酸化mRNA进行分析,分别鉴定出3.8 kb和4.0 kb的单一RNA种类,作为这两个物种中c-myb基因座的代表性转录本。因此,c-myb在人类和鸡中似乎是一个单一的基因座,在这两个远缘物种中具有相似的结构。我们制备的对DNA/RNA印迹具有更高灵敏度的特异性人类c-myb探针,应有助于对人类正常或肿瘤细胞或组织中的该基因进行分析。