Yoo O J, Yoon H S, Baek K H, Jeon C J, Miyamoto K, Ueno A, Agarwal K
Department of Biochemistry, Molecular Biology and Chemistry, University of Chicago, IL 60637.
Nucleic Acids Res. 1991 Mar 11;19(5):1073-9. doi: 10.1093/nar/19.5.1073.
The cDNA for the human elongation factor, TFIIS, has been cloned and expressed in E. coli with the T7 expression system. This 280-amino acid TFIIS protein is shorter by 21 residues than that of the mouse. The missing 21 residues are located in the amino-terminal region, which is not thought to be required for transcriptional stimulation. Apart from this gap, human and mouse proteins reveal 96% overall identity and 98.5% sequence similarity if conservative substitutions are taken into account. The bacterially expressed human protein and the purified calf thymus proteins are indistinguishable in their ability to stimulate transcript elongation by purified RNA polymerase II. Estimation of the native molecular size of the human protein in solution indicates that it exists as a dimer.
人类延伸因子TFIIS的cDNA已被克隆,并利用T7表达系统在大肠杆菌中表达。这种由280个氨基酸组成的TFIIS蛋白比小鼠的TFIIS蛋白短21个残基。缺失的21个残基位于氨基末端区域,一般认为该区域对转录刺激并非必需。除了这一差异外,若考虑保守性替换,人类和小鼠的蛋白在整体上有96%的同一性以及98.5%的序列相似性。细菌表达的人类蛋白与纯化的小牛胸腺蛋白在刺激纯化的RNA聚合酶II进行转录延伸的能力上并无差异。对溶液中人类蛋白天然分子大小的估计表明它以二聚体形式存在。