Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli
Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli
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DOI：
10.1007/s10969-006-9007-y
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发表时间：
2006-03-01
期刊：
影响因子：
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通讯作者：
Yokoyama, Shigeyuki
中科院分区：
文献类型：
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作者：
Chumpolkulwong, Namthip;Sakamoto, Kensaku;Yokoyama, Shigeyuki
我们分析了9个“稀有”密码子（阿加、AGG、AUA、CCC、CGA、CGG、CUA、GGA和UUA）对大肠杆菌偶联转录/翻译无细胞系统中基因表达的影响，并与基于细胞的表达系统进行了比较。每个报告基因含有一个罕见密码子的五个连续重复，或在某些实验中，三个连续重复。携带密码子CGA、CUA、GGA和UUA的基因的无细胞表达不受影响，尽管这些密码子（除了GGA）在E. coli细胞。其余5个密码子（阿加、AGG、AUA、CCC和CGG）的翻译在两个系统中都严重减少，并且在基于来自E.大肠杆菌细胞过量产生这些密码子的“次要”tRNA。
We analyzed the effect of nine 'rare' codons (AGA, AGG, AUA, CCC, CGA, CGG, CUA, GGA, and UUA) on gene expression in an Escherichia coli coupled transcription/translation cell-free system, in comparison with a cell-based expression system. Each reporter gene contained five consecutive repeats of a rare codon, or in some experiments, three consecutive repeats. The cell-free expression of the genes bearing the codons CGA, CUA, GGA, and UUA was not affected, although these codons, except for GGA, were inefficiently translated in E. coli cells. Translation of the remaining five codons (AGA, AGG, AUA, CCC, and CGG) was severely reduced in both systems, and was remarkably facilitated in the cell-free system based on an S30 extract from the E. coli cells overproducing 'minor' tRNAs for these codons.