Tymms M J, McInnes B
Centre for Molecular Biology and Medicine, Monash University, Clayton, Victoria, Australia.
Gene Anal Tech. 1988 Jan-Feb;5(1):9-15. doi: 10.1016/0735-0651(88)90021-0.
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-alpha 1 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-alpha l is equivalent to the yeast-synthesized protein. In contrast with the yeast-expression system, where some IFN-alpha analogs are poorly expressed, all analogs tested were well expressed in RRL.
我们研究了利用体外表达作为一种快速便捷的手段，来筛选通过体外诱变产生的大量干扰素（IFN）类似物。使用SP6 RNA聚合酶从DNA模板生成的IFN-α1 mRNA在兔网织红细胞裂解物（RRL）中能高效翻译。这种RRL合成的IFN-α1的抗病毒比活性与酵母合成的蛋白质相当。与酵母表达系统不同，在酵母表达系统中一些IFN-α类似物表达不佳，而在RRL中所有测试的类似物都表达良好。
