丙型肝炎病毒C区的DNA改组

DNA shuffling of hepatitis C virus C

目的:利用DNA改组技术进行不同基因型别丙型肝炎病毒(HCV)基因组C区的人工进化。方法:首先利用PCR扩增了三段具有较高序列同源性的460 bp基因片段,然后将其等量混合,在Mg2+存在的条件下,用脱氧核糖核酸酶(DNaseⅠ)切割成50 bp左右的小片段。这些小片段在不外加引物的条件下,利用PCR反应进行重聚,再将重聚物经过一轮正常的PCR扩增。结果:获得了与原片段大小相当的基因片段。结论:这一技术为进一步筛选高活性的HCV C区基因打下基础,有利于从一组序列同源性程度较高的基因库构建随机嵌合基因,并为改组其他基因家族提供了借鉴。

Objective ：To use DNA shuffling for directed evolution of HCV genotypes. Methods ：Three genes of 460 bp were obtained by PCR from genotype, respectively. After mixed with equimolar of each, these genes DNase Ⅰ into small fragments of about 50 bp under the existence of Mg^2＋. genome C genes with different three templates with different were further cut randomly by Results： These small fragments were successfully reassembled to form full genes with original size by one round of PCR without any external primers and another round of normal PCR amplification. Conclusion：This shuffling protocol has laid the foundation for further experiments in the screening for high activity of C gene. It may also help to construct chimera genes from a family of genes with high sequence homology and be used to evolve other gene families.