Jay E, Seth A K, Jay G
J Biol Chem. 1980 May 10;255(9):3809-12.
An icosadeoxyribonucleotide containing the several features found in prokaryotic mRNA ribosome binding sites has been synthesized. This sequence can stimulate the binding of initiator fMet-tRNAf to the ribosome to form a stable 71 S initiation complex identical with those induced by natural messengers. The binding of this synthetic ribosome binding site is absolutely dependent upon initiation factor IF3, and the bound fMet-tRNAf is sensitive to puromycin indicating the formation of a functional initiation complex. A heptadecadeoxyribonucleotide, identical with the icosanucleotide but lacking the terminal A-T-G codon, can also stimulate the stable binding of fMet-tRNAf to the ribosome, suggesting that the selection of the proper A-U-G initiation codon by fMet-tRNAf is subsequent to and a result of the recognition and binding of the fMet-tRNAf . 30 S ribosome complex to the initiation site. The prospect of ligating a similar synthetic ribosome binding site in front of a eukaryotic gene for cloning in an appropriate prokaryotic vector to assure the expresion of the protein is discussed.
一种含有原核生物信使核糖核酸（mRNA）核糖体结合位点中发现的几种特征的二十脱氧核糖核苷酸已被合成。该序列能刺激起始甲硫氨酰 - tRNAf与核糖体结合，形成与天然信使诱导形成的稳定71S起始复合物相同的复合物。这种合成核糖体结合位点的结合绝对依赖于起始因子IF3，并且结合的甲硫氨酰 - tRNAf对嘌呤霉素敏感，表明形成了功能性起始复合物。一种十七脱氧核糖核苷酸，与二十核苷酸相同但缺少末端A - T - G密码子，也能刺激甲硫氨酰 - tRNAf与核糖体的稳定结合，这表明甲硫氨酰 - tRNAf对合适的A - U - G起始密码子的选择是在甲硫氨酰 - tRNAf·30S核糖体复合物识别并结合到起始位点之后，并且是其结果。本文讨论了在真核基因前连接类似的合成核糖体结合位点以克隆到合适的原核载体中以确保蛋白质表达的前景。
