Specific transcription of an Acanthamoeba castellanii 5S RNA gene in homologous nuclear extracts.
Specific transcription of an Acanthamoeba castellanii 5S RNA gene in homologous nuclear extracts.
复制标题
同源核提取物中卡氏棘阿米巴 5S RNA 基因的特异性转录。
DOI：
10.1093/nar/19.7.1681
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发表时间：
1991
影响因子：
14.9
通讯作者：
Paule,MR
中科院分区：
文献类型：
--
作者：
Zwick,MG;Imboden,MA;Paule,MR
从卡氏变形虫（amoebacastellanii）中建立了RNA聚合酶III的体外转录系统。该系统依赖于克隆的5S RNA基因，并利用含有所有必需蛋白质组分的核提取物。该系统由完全同源的组件组装而成。引物延伸和RNA测序分析证实体外5S RNA转录物与从细胞中分离的5S RNA完全相同。转录复合物在5S RNA基因上异常迅速地形成，并且对于过量竞争模板的挑战是稳定的。构建了几个5′缺失突变体，表明−33的上游区域是β。缺失到+16表示转录所需的-33和+16之间的区域，这是典型的内部控制区域之外的区域。
An RNA polymerase IIIin vitrotranscription system has been developed from the protistAcanthamoeba castellanii. The system is dependent on a cloned 5S RNA gene and utilizes a nuclear extract which contains all the necessary protein components. The system is assembled from completely homologous components. Primer extension and RNA sequencing analysis confirm that thein vitro5S RNA transcript is identical to the 5S RNA isolated from cells. The transcription complex forms unusually rapidly on the 5S RNA gene and is stable to challenge by excess competitor templates. Several 5′ deletion mutants were constructed and indicate that the region upstream of −33 is dispensable. Deletion to +16 show the region between −33 and +16 to be required for transcription, a region outside the canonical internal control region.