A CONTINUOUS CELL-FREE TRANSLATION SYSTEM CAPABLE OF PRODUCING POLYPEPTIDES IN HIGH-YIELD
A CONTINUOUS CELL-FREE TRANSLATION SYSTEM CAPABLE OF PRODUCING POLYPEPTIDES IN HIGH-YIELD
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DOI：
10.1126/science.3055301
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发表时间：
1988-11-25
期刊：
影响因子：
56.9
通讯作者：
ALAKHOV, YB
中科院分区：
文献类型：
--
作者：
SPIRIN, AS;BARANOV, VI;ALAKHOV, YB
构建了一个无细胞翻译系统，该系统使用连续流动的进料缓冲液[包括氨基酸、三磷酸腺苷（ATP）和三磷酸鸟苷（GTP）]通过反应混合物，并连续去除多肽产物。该系统的原核生物（大肠杆菌）和真核生物（小麦胚胎，小麦）版本都已进行了测试。在这两种情况下，该系统已被证明具有长时间的活性，以高恒定速率合成多肽长达数十小时。以MS2噬菌体RNA和雀麦花叶病毒RNA 4为模板，在原核和真核系统中分别合成了每RNA 100个拷贝的病毒外壳蛋白，耗时20小时。用合成的降钙素信使RNA，在两种类型的连续翻译系统中，每个信使RNA产生150至300个拷贝的降钙素多肽，持续40小时。
A cell-free translation system has been constructed that uses a continuous flow of the feeding buffer [including amino acids, adenosine triphosphate (ATP), and guanosine triphosphate (GTP)] through the reaction mixture and a continuous removal of a polypeptide product. Both prokaryotic (Escherichia coli) and eukaryotic (wheat embryos, Triticum sp.) versions of the system have been tested. In both cases the system has proven active for long times, synthesizing polypeptides at a high constant rate for tens of hours. With the use of MS2 phage RNA or brome mosaic virus RNA 4 as templates, 100 copies of viral coat proteins per RNA were synthesized for 20 hours in the prokaryotic or eukaryotic system, respectively. With synthetic calcitonin messenger RNA, 150 to 300 copies of calcitonin polypeptide were produced per messenger RNA in both types of continuous translation systems for 40 hours.