利用非洲爪蟾卵母细胞提取物对硒代半胱氨酸tRNA[Ser]Sec基因进行体外转录分析。

Analysis of the selenocysteine tRNA[Ser]Sec gene transcription in vitro using Xenopus oocyte extracts.

作者信息

Park J M, Yang E S, Hatfield D L, Lee B J

机构信息

Laboratory of Molecular Genetics, Seoul National University, Korea.

出版信息

Biochem Biophys Res Commun. 1996 Sep 4;226(1):231-6. doi: 10.1006/bbrc.1996.1338.

DOI:10.1006/bbrc.1996.1338

PMID:8806619

Abstract

An in vitro transcription system was devised using a Xenopus oocyte S150 extract for analyzing expression of the Sec tRNA[Ser]Sec gene. The activator element, located at about -200 that is required for maximal expression of the tRNA[Ser]Sec gene in vivo, had no effect on tRNA[Ser]Sec transcription in vitro. In vitro transcription of this gene even tolerated mutations within or deletion of the PSE, showing marked contrast with the requirements observed in vivo. However, the TATA box was indispensable for basal level expression of the gene both in vivo and in vitro. The region spanning from the TATA box to the transcription start point (-33 to -1) was sufficient for Pol III to recognize the tRNA[Ser]Sec gene promoter in vitro.

摘要

设计了一种使用非洲爪蟾卵母细胞S150提取物的体外转录系统，用于分析Sec tRNA[Ser]Sec基因的表达。位于约-200处的激活元件是tRNA[Ser]Sec基因在体内最大表达所必需的，但对体外tRNA[Ser]Sec转录没有影响。该基因的体外转录甚至能耐受PSE内的突变或缺失，这与体内观察到的要求形成明显对比。然而，TATA框对于该基因在体内和体外的基础水平表达都是必不可少的。从TATA框到转录起始点（-33至-1）的区域足以使Pol III在体外识别tRNA[Ser]Sec基因启动子。