利用DNA改组技术创建新型传染性支气管炎病毒S1基因：对S1基因重组的影响
Using DNA shuffling to create novel infectious bronchitis virus S1 genes: implications for S1 gene recombination.
作者信息
Callison Scott, Hilt Deborah, Jackwood Mark
机构信息
Department of Avian Medicine, Poultry Diagnostic and Research Center, The University of Georgia, Athens, GA 30605, USA.
出版信息
Virus Genes. 2005 Aug;31(1):5-11. doi: 10.1007/s11262-004-2194-3.
We employed the staggered extension process (StEP) to shuffle the S1 genes from four infectious bronchitis virus (IBV) strains representing four unique serotypes. Upon creating a shuffled S1 gene library, we randomly selected 25 clones and analyzed them by DNA sequencing. In total, eleven clones contained novel S1 gene recombinants. Based on sequence data, each recombinant was unique and contained a full-length open reading frame. The average number of crossovers per recombinant was 5 and the average number of point mutations was 1.3, leading mostly to non-synonymous amino acid changes. No recombinant contained sequences from all four parental genes and no recombinant contained any sequence from the distantly related Delaware 072 strain. Our data suggests that recombination between distantly related IBV strains within the S1 gene probably does not readily occur. This finding is extremely important in light of the common industry vaccination practice of mixing different live-attenuated IBV strains.
我们采用交错延伸过程（StEP）对代表四种独特血清型的四种传染性支气管炎病毒（IBV）毒株的S1基因进行改组。构建改组后的S1基因文库后，我们随机挑选了25个克隆并通过DNA测序进行分析。总共11个克隆包含新型S1基因重组体。根据序列数据，每个重组体都是独特的，并且包含一个全长开放阅读框。每个重组体的交叉平均数量为5，点突变平均数量为1.3，主要导致非同义氨基酸变化。没有重组体包含来自所有四个亲本基因的序列，也没有重组体包含来自远缘相关的特拉华072毒株的任何序列。我们的数据表明，S1基因内远缘相关的IBV毒株之间的重组可能不容易发生。鉴于行业中混合不同活减毒IBV毒株的常见疫苗接种做法，这一发现极为重要。
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