Drummond D R, Armstrong J, Colman A
Nucleic Acids Res. 1985 Oct 25;13(20):7375-94. doi: 10.1093/nar/13.20.7375.
Synthetic RNAs coding for chicken lysozyme, calf preprochymosin and Xenopus globin were transcribed in vitro using Sp6 RNA polymerase. The effects of capping and adding a poly(A) tail on the stability, movement and translation of these RNAs in Xenopus oocytes was examined. Capping and polyadenylation increased stability of the transcripts, with at least 40% remaining intact 48 h after injection into oocytes. Capped poly(A)- transcripts moved more rapidly in oocytes than either capped poly(A)+ transcripts or naturally occurring mRNAs. The translational efficiency of most of the synthetic RNAs in oocytes increased with both capping and polyadenylation. The exception was one Xenopus globin transcript which had an unusual 3' end of 20As and 30Cs, where further polyadenylation decreased translational efficiency. Polyadenylation was essential for detectable expression of the synthetic RNAs in cultured cells, but decreased translation of the synthetic RNAs in vitro.
使用Sp6 RNA聚合酶在体外转录编码鸡溶菌酶、小牛前凝乳酶原和非洲爪蟾珠蛋白的合成RNA。研究了加帽和添加多聚腺苷酸尾对这些RNA在非洲爪蟾卵母细胞中的稳定性、移动性和翻译的影响。加帽和聚腺苷酸化增加了转录本的稳定性，注射到卵母细胞中48小时后至少有40%保持完整。加帽的多聚腺苷酸 - 转录本在卵母细胞中的移动速度比加帽的多聚腺苷酸 + 转录本或天然存在的mRNA都要快。大多数合成RNA在卵母细胞中的翻译效率随着加帽和聚腺苷酸化而提高。例外的是一种非洲爪蟾珠蛋白转录本，其3'端有20个A和30个C的不寻常序列，进一步的聚腺苷酸化会降低翻译效率。聚腺苷酸化对于合成RNA在培养细胞中的可检测表达至关重要，但会降低合成RNA在体外的翻译。
