Chang C C, Chen T T, Cox B W, Dawes G N, Stemmer W P, Punnonen J, Patten P A
Maxygen, Inc., 3410 Central Expressway, Santa Clara, CA 95051, USA.
Nat Biotechnol. 1999 Aug;17(8):793-7. doi: 10.1038/11737.
DNA shuffling of a family of over 20 human interferon-alpha (Hu-IFN-alpha) genes was used to derive variants with increased antiviral and antiproliferation activities in murine cells. A clone with 135,000-fold improved specific activity over Hu-IFN-alpha2a was obtained in the first cycle of shuffling. After a second cycle of selective shuffling, the most active clone was improved 285,000-fold relative to Hu-IFN-alpha2a and 185-fold relative to Hu-IFN-alpha1. Remarkably, the three most active clones were more active than the native murine IFN-alphas. These chimeras are derived from up to five parental genes but contained no random point mutations. These results demonstrate that diverse cytokine gene families can be used as starting material to rapidly evolve cytokines that are more active, or have superior selectivity profiles, than native cytokine genes.
通过对20多个人类α干扰素（Hu-IFN-α）基因家族进行DNA改组，在鼠细胞中获得了具有增强抗病毒和抗增殖活性的变体。在第一轮改组中获得了一个比Hu-IFN-α2a比活性提高了135,000倍的克隆。经过第二轮选择性改组后，活性最高的克隆相对于Hu-IFN-α2a提高了285,000倍，相对于Hu-IFN-α1提高了185倍。值得注意的是，三个活性最高的克隆比天然鼠α干扰素更具活性。这些嵌合体源自多达五个亲本基因，但没有随机点突变。这些结果表明，不同的细胞因子基因家族可以用作起始材料，以快速进化出比天然细胞因子基因更具活性或具有更优选择性的细胞因子。
