Hulen C, Henckès G, Legault-Démare J
Biochimie. 1977;59(2):179-88. doi: 10.1016/s0300-9084(77)80289-7.
Good evidence is provided that fMet-tRNA binding and aminoacid incorporation, when single-stranded DNA is used instead of mRNA in an E. coli cell-free system, are strictly dependent on the magnesium concentration. Ten sites homologous to the initiation sites of translation can be detected on denatured T5 stO DNA when using ribosomes and initiation factors from uninfected E. coli F. In S-30 extracts, at high magnesium concentrations and in the presence of neomycin, initiation of the translation of denatured T5 stO DNA begins anywhere on the molecule, and yet high molecular weight polypeptides are synthesized. The template potentiality of the denatured T5 stO DNA decreased when using ribosomes plus initiation factors and crude extracts from T5 stO-infected bacteria. By in vitro formation of initiation complexes sites analogous to initiation sites of translation were localized on T5 stO DNA molecules using single-stranded fragments separated by sedimentation in alkaline sucrose gradient.
有充分证据表明，在大肠杆菌无细胞系统中，当使用单链DNA代替mRNA时，甲酰甲硫氨酸 - tRNA结合和氨基酸掺入严格依赖于镁浓度。当使用来自未感染的大肠杆菌F的核糖体和起始因子时，在变性的T5 stO DNA上可以检测到十个与翻译起始位点同源的位点。在S - 30提取物中，在高镁浓度和新霉素存在下，变性T5 stO DNA的翻译起始在分子的任何位置开始，然而仍能合成高分子量多肽。当使用核糖体加起始因子以及来自T5 stO感染细菌的粗提取物时，变性T5 stO DNA的模板潜力降低。通过体外形成起始复合物，使用在碱性蔗糖梯度中沉降分离的单链片段，在T5 stO DNA分子上定位了类似于翻译起始位点的位点。
