Effect of estrogen on gene expression in the chick oviduct. In vitro transcription of the ovalbumin gene in chromatin.

Deoxyribonuclease I sensitivity of the nontranscribed sequences flanking the 5' and 3' ends of the ovomucoid gene and the ovalbumin and its related X and Y genes in hen oviduct nuclei

“…The ratio of unlabeled DNA to [3H]DNA was at least 107 in each analysis. Samples were denatured at 100 °C for 5 min and incubated at 68 °C for various time intervals ranging from 10 min to 40 h. Following hybridization, samples were treated with SI nuclease (Miles, 4800 units/vial) at 37 °C and nuclease resistant, trichloroacetic acid (CfiAcOH) precipitable radioactivity was determined as previously described (Harris et al, 1976). A contribution to SI nuclease counts resulting from reannealing of complementary strands of the nick-translated probes was monitored by preparation and parallel incubation of vials in which unlabeled chick driver DNA was replaced by an identical concentration of yeast RNA.…”

Regulation of gene expression in the chick oviduct. 18. Effect of estrogen on gene expression in the chick oviduct. Regulation of the ovomucoid gene

“…Samples were then treated with S| nuclease (4800 units) in 600 µL of 0.4 N NaCl, 0.2 N sodium acetate, 25 mM zinc acetate, pH 4.5, and the Si nuclease resistant hybrids were determined as described elsewhere (Harris et al, 1976).…”

“…These RNA excess hybridization experiments were performed in a final volume of 30 µ under conditions as described earlier (Harris et al, 1976). RNA, 4-5000 µg/mL, was boiled together with 0.14 ng of [3H]DNAom probe for 5 min.…”

“…RNA, 4-5000 µg/mL, was boiled together with 0.14 ng of [3H]DNAom probe for 5 min. The mixture was incubated at 68 °C for time intervals ranging from 15 min to 72 h. Following hybridization the samples were treated with Si nuclease (1600 units) and the Si nuclease resistant hybrids were determined as previously described (Harris et al, 1976).…”

Effect of estrogen on ovalbumin gene expression in differentiated nontarget tissues

“…Samples were then incubated at 68 °C for time intervals ranging from 10 min to 70 h. Following hybridization, the samples were treated with St nuclease (1600 units), and the S! nuclease resistant hybrids were determined as previously described (Harris et al, 1976). Equivalent Rat values, obtained at 0.18 M NaCl, were plotted (Britten et al, 1974), and R0fi/2 was determined at half-maximum of hybridization.…”