Colbert D A, Knoll B J, Woo S L, Mace M L, Tsai M J, O'Malley B W
Biochemistry. 1980 Nov 25;19(24):5586-92. doi: 10.1021/bi00565a020.
We describe the isolation of recombinant phages from a chicken gene library which contain two genes designated X and Y. These two genes are linked to the ovalbumin gene (OV) in the order 5'-X-Y-ovalbumin-3' [Royal, A., Garapin, A., Cami, B., Perrin, F., Mandel, J. L., LeMeur, M., Bregegegre, F., Gannon, F, LePennec, J. P., Chambon, P., & Kourilsky, P. (1979) Nature (London) 279, 125-132]. Both genes contain multiple intervening sequences and share limited sequence homology with the authentic ovalbumin gene but are expressed in oviduct cells at different levels. X and Y hybridization probes were prepared in order to study the expression and the relative hormonal responsiveness of these three genes in chicken oviduct. The sequence specificity of the probes was demonstrated by Southern hybridization assays. Northern hybridization studies using the X and Y gene probes indicated the presence of putative precursor molecules in stimulated oviduct ribonucleic acid preparations, which differ in size from those observed for ovalbumin. R0t analysis has demonstrated that, similar to the ovalbumin gene, the level of X and Y gene transcripts is increased by the steroid hormone estrogen, but to varying degrees. The extent of hormonal responsiveness of the three closely related genes is in the order (normalized) relative to ovalbumin of OV:Y:X congruent to 100:10:1. Pulse-labeling studies of these three closely linked genes suggest that in estrogen-stimulated oviduct, the markedly different steady-state levels of the X, Y, and ovalbumin gene transcripts reflect their differential transcription rates.
我们描述了从鸡基因文库中分离出重组噬菌体,这些噬菌体含有两个命名为X和Y的基因。这两个基因与卵清蛋白基因(OV)相连,顺序为5'-X-Y-卵清蛋白-3' [Royal, A., Garapin, A., Cami, B., Perrin, F., Mandel, J. L., LeMeur, M., Bregegegre, F., Gannon, F, LePennec, J. P., Chambon, P., & Kourilsky, P. (1979) 《自然》(伦敦)279, 125 - 132]。这两个基因都含有多个间隔序列,与真正的卵清蛋白基因有有限的序列同源性,但在输卵管细胞中以不同水平表达。制备了X和Y杂交探针,以研究这三个基因在鸡输卵管中的表达及相对激素反应性。通过Southern杂交分析证明了探针的序列特异性。使用X和Y基因探针的Northern杂交研究表明,在受刺激的输卵管核糖核酸制剂中存在推定的前体分子,其大小与卵清蛋白的不同。R0t分析表明,与卵清蛋白基因相似,X和Y基因转录本的水平会被类固醇激素雌激素提高,但程度不同。这三个密切相关基因的激素反应程度顺序为(相对于卵清蛋白归一化)OV:Y:X约为100:10:1。对这三个紧密连锁基因的脉冲标记研究表明,在雌激素刺激的输卵管中,X、Y和卵清蛋白基因转录本明显不同的稳态水平反映了它们不同的转录速率。