胶原蛋白信使核糖核酸的鉴定。
The identification of collagen messenger RNA.
作者信息
Boedtker H, Crkvenjakov R B, Last J A, Doty P
出版信息
Proc Natl Acad Sci U S A. 1974 Oct;71(10):4208-12. doi: 10.1073/pnas.71.10.4208.
RNA isolated from calvaria of 16- to 18- day-old chick embryos, assayed in rabbit reticulocyte lysates, programs the synthesis of a collagenase-sensitive protein with the molecular weight of collagen pro-alpha-chains. When RNA labeled with [(3)H]uridine for 2 hr and chased for 1 or 2 hr was electrophoresed on aqueous polyacrylamide gels, most of the radioactivity not in 28S or 18S rRNA migrated with an apparent molecular weight of about 1,800,000. After oligo(dT)-cellulose chromotography and analysis in 99% formamide gels, this nonribosomal, rapidly labeled calvaria RNA species migrates at 28S-30S and thus has a molecular weight of at least 1,600,000. Both the ability to program the synthesis of collagenase-sensitive protein in reticulocyte lysates and the presence of a single prominent rapidly labeled 30S peak in acrylamide gels strongly support the deduction that there is only one major mRNA species in calvaria and that this species is collagen messenger RNA.
从16至18日龄鸡胚的颅骨中分离出的RNA，在兔网织红细胞裂解物中进行检测，可指导合成一种对胶原酶敏感的蛋白质，其分子量与胶原蛋白前α链相同。当用[³H]尿苷标记2小时并追踪1或2小时后的RNA在水性聚丙烯酰胺凝胶上进行电泳时，大部分不在28S或18S rRNA中的放射性物质迁移时的表观分子量约为1,800,000。经过寡聚（dT）-纤维素层析并在99%甲酰胺凝胶中分析后，这种非核糖体的、快速标记的颅骨RNA物种在28S - 30S处迁移，因此分子量至少为1,600,000。在网织红细胞裂解物中指导合成对胶原酶敏感的蛋白质的能力以及在丙烯酰胺凝胶中存在一个单一突出的快速标记的30S峰，都有力地支持了这样的推断：颅骨中只有一种主要的mRNA物种，且该物种是胶原蛋白信使RNA。
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