利用SP6聚合酶和兔网织细胞裂解液高效体外表达干扰素α类似物

Martin J. Tymms, Beth McInnes
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引用次数: 25

摘要

我们已经研究了使用体外表达作为筛选使用体外诱变产生的大量干扰素(IFN)类似物的快速和方便的手段。使用SP6 RNA聚合酶从DNA模板产生的IFN-α1mRNA在兔网织红细胞裂解物(RRL)中被有效翻译。该RRL合成的IFN-α1的抗病毒特异性活性相当于酵母合成的蛋白质。与酵母表达系统相反,在酵母表达系统中,一些IFN-α类似物表达不佳,所有测试的类似物在RRL中都表达良好。
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Efficient in vitro expression of interferon α analogs using SP6 polymerase and rabbit reticulocyte lysate

We have investigated the use of in vitro expression as a quick and convenient means of screening large numbers of interferon (IFN) analogs generated using in vitro mutagenesis. The IFN-α1 mRNA generated from DNA template using SP6 RNA polymerase is efficiently translated in rabbit reticulocyte lysate (RRL). The antiviral specific activity of this RRL-synthesized IFN-α1 is equivalent to the yeast-synthesized protein. In contrast with the yeast-expression system, where some IFN-α analogs are poorly expressed, all analogs tested were well expressed in RRL.

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Gene targeting in murine embryonic stem cells: Introduction of specific alterations into the mammalian genome A solution hybridization method for quantification of mRNAs: Determining the amount and stability of oncogene mRNA The use of transgenic mice for short-term, in vivo mutagenicity testing Author index volume 7 Subject index volume 7
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