Liu C P, Slate D L, Gravel R, Ruddle F H
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4503-6. doi: 10.1073/pnas.76.9.4503.
We have used direct microinjection of messenger RNA into individual mouse and human cells to assay for specific translation products. We have been able to detect the synthesis of human fibroblast interferon, thymidine, kinase, hypoxanthine phosphoribosyltransferase, adenine phosphoribosyltransferase, and propionyl-CoA carboxylase in response to injected mRNA. Using the interferon system as a model, we have quantitated interferon synthesis and followed partial purification of interferon mRNA sequences on sucrose density gradients. The methods we have utilized should be applicable to other systems in which sensitive assays exist for gene products and should provide a screening procedure for isolating specific mRNA sequences.
我们已将信使核糖核酸直接显微注射到单个小鼠和人类细胞中,以检测特定的翻译产物。我们能够检测到,响应注射的信使核糖核酸,人类成纤维细胞干扰素、胸苷激酶、次黄嘌呤磷酸核糖转移酶、腺嘌呤磷酸核糖转移酶和丙酰辅酶A羧化酶的合成。以干扰素系统作为模型,我们对干扰素的合成进行了定量,并在蔗糖密度梯度上对干扰素信使核糖核酸序列进行了部分纯化。我们所采用的方法应适用于对基因产物存在灵敏检测方法的其他系统,并应为分离特定信使核糖核酸序列提供一种筛选程序。