Lund E, Dahlberg J E
Department of Physiological Chemistry, University of Wisconsin-Madison 53706.
EMBO J. 1989 Jan;8(1):287-92. doi: 10.1002/j.1460-2075.1989.tb03375.x.
We have developed a DNA-dependent in vitro transcription system for vertebrate snRNA genes. By isolating the nuclei (germinal vesicles, GVs) of Xenopus laevis oocytes under oil to maintain the in vivo composition of their internal milieu, we are able to prepare nuclei that retain their ability to synthesize snRNAs efficiently. Homogenates of these GVs synthesize correctly initiated and terminated U1 snRNA using exogenous X.laevis U1 genes as templates. The templates may be either injected into the nucleus prior to its isolation or added to the nuclear homogenate.
我们已经开发出一种用于脊椎动物小核RNA（snRNA）基因的依赖DNA的体外转录系统。通过在油下分离非洲爪蟾卵母细胞的细胞核（生发泡，GVs）以维持其内部环境的体内组成，我们能够制备出保留高效合成snRNAs能力的细胞核。这些GVs的匀浆物以外源非洲爪蟾U1基因作为模板，正确起始和终止合成U1 snRNA。模板既可以在细胞核分离之前注射到细胞核中，也可以添加到细胞核匀浆中。
