在经雌二醇处理的雏鸡肝脏中鉴定卵黄蛋白原mRNA的一种大的前体。
Identification of a large precursor of vitellogenin mRNA in the liver of estradiol-treated chicks.
作者信息
Jost J P, Pehling G, Ohno T, Cozens P
出版信息
Nucleic Acids Res. 1978 Dec;5(12):4781-93. doi: 10.1093/nar/5.12.4781.
Studies were performed to determine whether vitellogenin mRNA from avian liver has a precursor molecule or not. Total cellular RNA was prepared from estradiol-treated chicken liver in the presence of 8 M guanidine HCl, 2-mercaptoethanol and aurintricarboxylic acid. After denaturation, RNA was fractionated on sodium dodecylsulfate-sucrose gradients and large size RNA was analyzed under stringent conditions on 85% formamide-sucrose gradients at 25 degrees C. RNA fractions collected from the gradients were hybridized with vitellogenin (3H)-cDNA. Besides mature vitellogenin mRNA (32S, 7,000 nucleotides) vitellogenin sequences were also found in RNA fractions ranging from 38-50S with a peak at 45-50S (12-15,000 nucleotides). Only 5-10% of the putative 38-50S pmRNA is polyadenylated. We calculated that the half-life of vitellogenin pmRNA is about 3-4 minutes. We conclude that vitellogenin mRNA has a precursor which is twice the size of the mature mRNA.
开展了多项研究以确定来自鸟类肝脏的卵黄蛋白原信使核糖核酸（mRNA）是否具有前体分子。在8M盐酸胍、2-巯基乙醇和金精三羧酸存在的情况下，从经雌二醇处理的鸡肝脏中制备总细胞核糖核酸（RNA）。变性后，RNA在十二烷基硫酸钠-蔗糖梯度上进行分级分离，大尺寸RNA在25摄氏度下于85%甲酰胺-蔗糖梯度的严格条件下进行分析。从梯度中收集的RNA级分与卵黄蛋白原（3H）-互补脱氧核糖核酸（cDNA）杂交。除了成熟的卵黄蛋白原mRNA（32S，7000个核苷酸）外，在38 - 50S的RNA级分中也发现了卵黄蛋白原序列，峰值在45 - 50S（12000 - 15000个核苷酸）。推测的38 - 50S前体mRNA（pmRNA）中只有5 - 10%是聚腺苷酸化的。我们计算出卵黄蛋白原pmRNA的半衰期约为3 - 4分钟。我们得出结论，卵黄蛋白原mRNA有一个前体，其大小是成熟mRNA的两倍。
Zotero 插件