Cloning and characterization of the TATA-binding protein of the silkworm Bombyx mori.
Cloning and characterization of the TATA-binding protein of the silkworm Bombyx mori.
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DOI：
10.1016/s0378-1119(98)00460-0
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发表时间：
1998-10
期刊：
影响因子：
3.5
通讯作者：
C. Ouyang;K. U. Sprague
中科院分区：
文献类型：
--
作者：
C. Ouyang;K. U. Sprague
TATA结合蛋白是所有三种真核细胞核RNA聚合酶所需的通用转录因子。为了研究TBP在家蚕tRNA基因转录中的作用，我们从家蚕cDNA文库中克隆了TBP cDNA。与大多数其他真核生物一样，家蚕中的TBP由单拷贝基因编码，并含有高度保守的C-末端结构域，包括一个碱性区域和两个直接重复序列。在保守性较低的N-末端结构域中，家蚕TBP表现出一些特征，如富含谷氨酰胺的延伸和三个不完美的Pro-Met-Thr样重复序列，这些特征也在果蝇和人TBP中发现。家蚕TBP在大肠杆菌中表达并纯化至表观均一性，以纳摩尔亲和力结合野生型腺病毒主要晚期启动子的TATA元件。
The TATA-binding protein is a general transcription factor required by all three eukaryotic nuclear RNA polymerases. In order to study the function of this protein in the transcription of tRNA genes in the silkworm Bombyx mori, we have cloned TBP cDNA from a silkworm cDNA library. As in most other eukaryotes, TBP in silkworms is encoded by a single copy gene and contains a highly conserved C-terminal domain that includes a basic region and two direct repeats. In the less conserved N-terminal domain, silkworm TBP exhibits characteristics such as a glutamine-rich stretch and three imperfect Pro-Met-Thr-like repeats that are also found in Drosophila and human TBP. Silkworm TBP expressed in Escherichia coli and purified to apparent homogeneity binds the TATA element of the wild-type adenovirus major late promoter with nanomolar affinity.