A tRNA gene of Xenopus laevis contains at least two sites promoting transcription.
A tRNA gene of Xenopus laevis contains at least two sites promoting transcription.
复制标题
非洲爪蟾的 tRNA 基因包含至少两个促进转录的位点。
DOI：
10.1093/nar/7.7.1749
复制
发表时间：
1979
影响因子：
14.9
通讯作者：
M. Birnstiel
中科院分区：
文献类型：
--
作者：
A. Kressmann;H. Hofstetter;E. Di Capua;R. Grosschedl;M. Birnstiel
一小段克隆的DNA片段被Hae III酶切成前部和后部，该片段包含非洲爪哇tRNA1met基因表达的所有遗传信息。利用EcoRI连接子将这两个限制性片段克隆到PCR1中。从这些tDNA亚克隆出发，构建了一系列新的重组体。通过体外转录系统和卵母细胞注射技术检测克隆DNA的转录活性。结果表明，克隆的基因单位的5‘端和3’端都是转录所必需的。我们已经建立了tRNA1met基因的功能组织模型。我们认为该基因在基因单元的3‘端附近含有一个调控位点。为了在体内和体外进行转录，需要在基因的5‘端附近有一个特定的起始点。
A small cloned DNA segment previously shown to contain all genetic information for the expression of the tRNA1met gene of Xenopus laevis was cleaved into an anterior and posterior portion by Hae III restriction. Both restriction fragments were cloned in pCR1 using EcoRI linkers. Starting from these tDNA subclones, a series of new recombinants were constructed. The transcriptional activity of the cloned DNA's was tested both in an in vitro transcriptional system and by means of the oocyte injection technique. It was shown that both the 5' and 3' ends of the cloned gene unit were essential for transcription. We have developed a model for the functional organization of the tRNA1met gene. We propose that the gene contains a regulatory site situated near the 3' portion of the gene unit. For transcription to occur both in vivo and in vitro a specific initiation site near the 5' end of the gene is required.