METHOD FOR CONSTRUCTING RECOMBINANT BACTERIUM FOR PRODUCING NON-NATIVE PROTEIN, AND UTILIZATION OF SAME
申请人:Riken
公开号:EP2584037A1
公开(公告)日:2013-04-24
The present invention provides a novel method of producing a recombinant bacterium for production of a non-natural protein, including: (1) expressing tRNA in a bacterium, which tRNA recognizes UAG codon; (2) expressing an aminoacyl-tRNA synthetase in the bacterium, which aminoacyl-tRNA synthetase acylates the tRNA with a non-natural amino acid or an α-hydroxy acid; (3) (i) introducing a DNA construct into the bacterium, which DNA construct is for expressing, in the absence of a release factor for terminating translation at UAG codon, a function of at least one gene selected from the group consisting of genes each of which loses its function when a gene that codes for the release factor is defective and/or introducing an alteration into said at least one gene in a chromosome of the bacterium, which alteration is for expressing the function of said at least one gene in the absence of the release factor; and (4) causing the gene that codes for the release factor in the bacterium to be defective.
本发明提供了一种生产非天然蛋白质的重组细菌的新方法,包括:(1) 在细菌中表达 tRNA,该 tRNA 识别 UAG 密码子;(2) 在细菌中表达氨基酰-tRNA 合成酶,该氨基酰-tRNA 合成酶用非天然氨基酸或 α-羟基酸酰化 tRNA;(3) (i) 向细菌中导入 DNA 构建体,该 DNA 构建体用于在没有用于在 UAG 密码子处终止翻译的释放因子的情况下表达至少一个基因的功能,该基因选自以下基因组:当编码释放因子的基因有缺陷时,每个基因都会失去其功能;和/或在细菌的染色体中向所述至少一个基因导入改变,该改变用于在没有释放因子的情况下表达所述至少一个基因的功能;(4) 使细菌中编码释放因子的基因出现缺陷。