Nicole L M, Tanguay R M
Ontogénèse et Génétique Molécularies, Centre Hospitalier Université, Laval. Ste-Foy, Québec, Canada.
Biosci Rep. 1987 Mar;7(3):239-46. doi: 10.1007/BF01124795.
The specificity of action of antisense RNA for one of Drosophila low molecular weight heat shock proteins (hsp 23) was tested at the translational level using the rabbit reticulocyte lysate cell-free system. T7 polymerase-driven transcripts of hsp 23 in the antisense orientation were mixed with mRNA from heat-shocked cells under various stringency conditions prior to translation in vitro. Although the four small hsps show considerable sequence homology in their coding sequences, antisense hsp 23 RNA was shown to specifically inhibit hsp 23 mRNA translation under both high (formamide, 45 degrees C) and low stringency (37 degrees C) conditions. This suggests that the 5' leader and the ribosome binding region of mRNA are of prime importance in the specificity of action of antisense RNA at the translational level.
利用兔网织红细胞裂解物无细胞系统，在翻译水平测试了反义RNA对果蝇一种低分子量热休克蛋白（hsp 23）的作用特异性。在体外翻译之前，将T7聚合酶驱动的反义方向的hsp 23转录本与热休克细胞的mRNA在各种严格条件下混合。尽管这四种小热休克蛋白在编码序列上显示出相当程度的序列同源性，但反义hsp 23 RNA在高严格条件（甲酰胺，45℃）和低严格条件（37℃）下均能特异性抑制hsp 23 mRNA的翻译。这表明mRNA的5'前导序列和核糖体结合区域在反义RNA翻译水平的作用特异性中至关重要。
