EFFICIENCY OF THE 5'-TERMINAL SEQUENCE (OMEGA) OF TOBACCO MOSAIC-VIRUS RNA FOR THE INITIATION OF EUKARYOTIC GENE TRANSLATION IN ESCHERICHIA-COLI
EFFICIENCY OF THE 5'-TERMINAL SEQUENCE (OMEGA) OF TOBACCO MOSAIC-VIRUS RNA FOR THE INITIATION OF EUKARYOTIC GENE TRANSLATION IN ESCHERICHIA-COLI
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DOI：
10.1111/j.1432-1033.1992.tb17271.x
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发表时间：
1992-10-01
期刊：
影响因子：
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通讯作者：
ABOUHAIDAR, MG
中科院分区：
文献类型：
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作者：
IVANOV, IG;ALEXANDROVA, R;ABOUHAIDAR, MG
最近的研究表明，烟草花叶病毒RNA的5'前导序列（OMEGA序列）在原核生物和真核生物中对mRNA的翻译都有一定的促进作用。为了评估OMEGA启动大肠杆菌中mRNA翻译的效率，与Shine-Dalgarno（S/D）序列相比，我们将8个不同的真核基因插入到两种类型的大肠杆菌中。含有一个组成型启动子（P1）但具有不同的启动子起始位点（分别为S/D或OMEGADELTA 3序列）的大肠杆菌表达载体。通过测量蛋白质的产量以及mRNA的水平和稳定性来评估这些载体在体内的转录和翻译效率。我们报道，OMEGADELTA 3取代S/D使表达蛋白的产量降低4 - 1900倍，基因特异性mRNA的含量降低约7倍。然而，与S/D序列相比，在OMEGA Δ 3的翻译控制下表达的蛋白质水平对5 ′编码区的变化不太敏感。我们还报告称，OMEGA序列包含一个与大肠杆菌小核糖体亚基RNA（rRNA）互补的10 - 12个核苷酸区域。coli、Eikenella corrodens和非洲爪蟾（Xenopus laevis），以及水稻（Oryza sativa）的（小核糖体）亚基的rRNA。
Recent studies have demonstrated that the 5' leader (OMEGA sequence) of tobacco mosaic virus RNA has a certain enhancing capacity for translation of mRNA in both prokaryotes and eukaryotes. In order to estimate the efficiency of OMEGA to initiate translation of mRNA in Escherichia coli, in comparison to the Shine-Dalgarno (S/D) sequence, we have inserted eight different eukaryotic genes into two types of E. coli expression vectors containing one constitutive promoter (P1) but different translation-initiation sites (S/D or OMEGADELTA3 sequence, respectively). The efficiency of transcription and translation in vivo was evaluated for these vectors by measuring the yield of protein and both the level and stability of MRNA. We report that substitution of OMEGADELTA3 for S/D decreases the yield of expressed protein 4 - 1900-fold and the content of gene-specific mRNA is decreased by about sevenfold. However, in comparison with the S/D sequence, the level of protein expressed under the translational control of OMEGADELTA3 is less sensitive to changes in the 5' coding region. We also report that the OMEGA sequence contains a region of 10 - 12 nucleotides complementary to the small ribosomal subunit RNA (rRNA) of E. coli, Eikenella corrodens and Xenopus laevis, and to the rRNA of the (small ribosomal) subunit of Oryza sativa.