Groudine M, Peretz M, Weintraub H
Division of Genetics, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Mol Cell Biol. 1981 Mar;1(3):281-8. doi: 10.1128/mcb.1.3.281-288.1981.
We used recombinant chicken deoxyribonucleic acid clones containing embryonic and adult beta-globin genes and "runoff" endogenous nuclear transcription to investigate the expression of embryonic and adult beta-globin genes during hematopoiesis in the developing chicken embryo. Purified, cloned deoxyribonucleic acids were digested with various restriction enzymes, separated on agarose gels, blotted to nitrocellulose, and annealed with purified nuclear [32P]ribonucleic acid synthesized in vitro from embryonic or adult red cell nuclei. Transcription of the respective globin genes was assayed by hybridization of nuclear [32P]ribonucleic acid to specific restriction fragments containing adult or embryonic coding sequences. Our results indicate that little, if any, transcription from the adult or embryonic beta-globin genes is detectable in the heterologous red cell nuclei, even under conditions in which ribonucleic acid processing probably does not occur.
我们使用了含有胚胎和成体β-珠蛋白基因的重组鸡脱氧核糖核酸克隆以及“连续”内源性核转录，来研究发育中的鸡胚造血过程中胚胎和成体β-珠蛋白基因的表达。将纯化的克隆脱氧核糖核酸用各种限制性内切酶消化，在琼脂糖凝胶上分离，印迹到硝酸纤维素膜上，并用从胚胎或成体红细胞核体外合成的纯化核[32P]核糖核酸进行退火处理。通过将核[32P]核糖核酸与含有成体或胚胎编码序列的特定限制性片段杂交，来检测各自珠蛋白基因的转录。我们的结果表明，即使在可能不发生核糖核酸加工的条件下，在异源红细胞核中也几乎检测不到来自成体或胚胎β-珠蛋白基因的转录。
