嗜热栖热菌多肽释放因子1的克隆与过表达
Cloning and overexpression of polypeptide release factor 1 of Thermus thermophilus.
作者信息
Ito K, Nakamura Y
机构信息
Department of Tumor Biology, Institute of Medical Science, University of Tokyo, Japan.
出版信息
Biochimie. 1997 May;79(5):287-92. doi: 10.1016/s0300-9084(97)83516-x.
PMID:9258437
Abstract
A prfA gene encoding polypeptide release factor RF1 was cloned from Thermus thermophilus. T thermophilus RF1 shares 68% homology with Escherichia coli RF1, and its overproduction reduced readthrough translation of UAG, not of UGA, in the lacZ gene. Rapid purification of T thermophilus RF1 was achieved by T7-RNA polymerase driven overexpression of T thermophilus RF1 protein with a C-terminal histidine tag.
摘要
从嗜热栖热菌中克隆了一个编码多肽释放因子RF1的prfA基因。嗜热栖热菌RF1与大肠杆菌RF1具有68%的同源性，其过量表达降低了lacZ基因中UAG而非UGA的通读翻译。通过T7 - RNA聚合酶驱动带有C端组氨酸标签的嗜热栖热菌RF1蛋白的过量表达，实现了嗜热栖热菌RF1的快速纯化。
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