Release factor one is nonessential in Escherichia coli.

Release factor one is nonessential in Escherichia coli.
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DOI:
10.1021/cb300229q
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发表时间:
2012-08-17
影响因子:
4
通讯作者:
Wang, Lei
Wang, Lei
中科院分区:
生物学2区
文献类型:
--
作者:
Johnson, David B. F.;Wang, Chong;Xu, Jianfeng;Schultz, Matthew D.;Schmitz, Robert J.;Ecker, Joseph R.;Wang, Lei

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将终止密码子重新编码为氨基酸可能会为生物合成新的蛋白质和生物体特性提供正交的遗传系统。虽然在现存的生物体中已经发现了终止密码子的重新分配,但还缺乏一个模式生物来研究重新分配过程和指导密码的进化。释放因子(RFs)阻止了终止密码子的完全重新分配,该因子识别终止密码子以终止翻译。在这里,我们发现RF1可以无条件地从各种大肠杆菌菌株中剔除,这表明据报道必不可少的RF1对于大肠杆菌物种来说通常是必不可少的。发现RF1的明显重要性是因为突变体RF2在终止所有UAA终止密码子方面效率低下;野生型RF2足以敲除RF1。RF1基因敲除菌株是自主的,并毫不含糊地重新分配UAG,在多个位点编码天然或非天然氨基酸(UAA),为研究密码进化提供了以前不可用的模型,并为利用UAA进化新的生物功能提供了独特的宿主。
Recoding a stop codon to an amino acid may afford orthogonal genetic systems for biosynthesizing new protein and organism properties. Although reassignment of stop codons has been found in extant organisms, a model organism is lacking to investigate the reassignment process and to direct code evolution. Complete reassignment of a stop codon is precluded by release factors (RFs), which recognize stop codons to terminate translation. Here we discovered that RF1 could be unconditionally knocked out from various Escherichia coli stains, demonstrating that the reportedly essential RF1 is generally dispensable for the E. coli species. The apparent essentiality of RF1 was found to be caused by the inefficiency of a mutant RF2 in terminating all UAA stop codons; a wild type RF2 was sufficient for RF1 knockout. The RF1-knockout strains were autonomous and unambiguously reassigned UAG to encode natural or unnatural amino acids (Uaas) at multiple sites, affording a previously unavailable model for studying code evolution and a unique host for exploiting Uaas to evolve new biological functions.
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