Morris G F, Price D H, Marzluff W F
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3674-8. doi: 10.1073/pnas.83.11.3674.
A soluble extract prepared from blastula nuclei of sea urchin (Lytechinus variegatus) embryos accurately transcribes cloned sea urchin DNA. This extract synthesizes U1 RNA using a cloned U1 RNA gene as a template. The U1 RNA is initiated accurately, and a portion of the transcripts has the correct 3' end as judged by gel electrophoresis. Longer transcripts also are formed that extend at least 280 bases 3' of the gene, and some extend as far as 800 bases 3' of the gene. A template containing 203 bases 5' of the gene gave as efficient transcription as did the whole U1 gene. Accurate 3' end formation was obtained with a template extending only 34 bases 3' of the gene, but efficient 3' end formation required sequences between 34 and 67 bases 3' of the gene.
从海胆(Lytechinus variegatus)胚胎囊胚细胞核制备的可溶性提取物能精确转录克隆的海胆DNA。该提取物以克隆的U1 RNA基因为模板合成U1 RNA。U1 RNA起始准确,通过凝胶电泳判断,一部分转录本具有正确的3'末端。还形成了更长的转录本,其在基因的3'端至少延伸280个碱基,有些延伸至基因的3'端达800个碱基。基因5'端包含203个碱基的模板与整个U1基因一样能高效转录。仅在基因3'端延伸34个碱基的模板能实现准确的3'末端形成,但高效的3'末端形成需要基因3'端34至67个碱基之间的序列。