Transcription in nuclei isolated from carrot protoplasts: effects of exogenous DNA
Transcription in nuclei isolated from carrot protoplasts: effects of exogenous DNA
复制标题
从胡萝卜原生质体分离的细胞核中的转录：外源 DNA 的影响
DOI：
10.1016/0014-5793(82)80284-6
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发表时间：
1982
期刊：
影响因子：
3.5
通讯作者：
P. Lurquin
中科院分区：
文献类型：
--
作者：
R. Calza;Suzanne M. Oelke;P. Lurquin
各种类型的分离的植物核在体外合成RNA [1 -8]。外源提供的DNA可以在分离的植物细胞核中转录，RNA刺激合成[7，8]。这表明外源DNA被植物原生质体吸收后可能在体内转录。此外，脂质体介导的胡萝卜原生质体的DNA摄取已被证明[9，13]，发现供体DNA的一部分与受体细胞核相关。因此，我们的目标是建立一个简单的植物体外转录系统，使我们能够确定植物RNA聚合酶II是否能够实现有意义的转录克隆的原核和真核基因。由于我们的兴趣是基因工程胡萝卜原生质体，我们已经确定了条件下，分离的胡萝卜核积极合成RNA。我们还表明，外源DNA刺激体外RNA合成，如181。这种刺激可能部分是由于抑制了核制剂中存在的核糖核酸酶。
Various types of isolated plant nuclei synthesize RNA in vitro [l-8]. Exogenously supplied DNA could be transcribed and RNA stimulated synthesis in isolated plant nuclei [7, 8]. This suggests that foreign DNA taken up by, eg, plant protoplasts might be transcribed in vivo. Furthermore, liposome-mediated DNA uptake by carrot protoplasts has been demonstrated[9, 13], part of the donor DNA being found associated with recipient nuclei. Thus, our goal is to develop a simple plant in vitro transcription system allowing-us to determine whether plant RNA polymerase II is able to achieve meaningful transcription of cloned prokaryotic and eukaryotic genes.In vitro RNA synthesis in carrot nuclei has not yet been described. Since our interest is to genetically engineer carrot protoplasts, we have determined conditions under which isolated carrot nuclei actively synthesized RNA. We also show that exogenous DNA stimulated in vitro RNA synthesis as observed in 181. Such stimulation could be due in part to the inhibition of ribonuclease present in the nuclei preparations.