FUNCTIONAL MESSENGER-RNAS ARE PRODUCED BY SP6 INVITRO TRANSCRIPTION OF CLONED CDNAS
FUNCTIONAL MESSENGER-RNAS ARE PRODUCED BY SP6 INVITRO TRANSCRIPTION OF CLONED CDNAS
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DOI：
10.1093/nar/12.18.7057
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发表时间：
1984-01-01
影响因子：
14.9
通讯作者：
MELTON, DA
中科院分区：
文献类型：
--
作者：
KRIEG, PA;MELTON, DA
我们描述了一种合成微克量的真核信使 RNA 的方法。注射到青蛙卵母细胞的细胞质中并添加到麦芽提取物中表明这些合成的 RNA 可以有效地发挥信使 RNA 的作用。我们证实 mRNA 上的 5' 帽对于注射卵母细胞中的翻译至关重要，并表明大多数 3' 侧翼区域（包括多聚 A 尾）可以被删除，而无需废除蛋白质合成。 mRNA 合成方法涉及已克隆到 SP6 载体中的 cDNA 的体外转录（在随附论文中描述）。这种方法使人们能够从任何 cDNA 克隆中产生大量的 mRNA 和蛋白质。
We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5′ cap on the mRNA is essential for translation in injected oocytes and show that most of the 3′ flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involvesinvitrotranscription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.