Asia1型口蹄疫病毒感染性克隆的建立

Construction of infectious clone for Asia1 foot-and-mouth disease virus

为构建Asia1型口蹄疫病毒Asia1/YS/CHA/05株的感染性克隆,根据该毒株的全基因组序列,将其分为6段进行RT-PCR扩增,并拼接克隆至pBluescript II SK(+)载体中,构建了全长基因组的cDNA克隆pAsi。用NotⅠ将质粒线性化后体外转录并转染BHK-21细胞,转染细胞中出现明显的细胞病变。RT-PCR结合序列分析表明,拯救病毒中作为人工设计的分子标记PstⅠ酶切位点被消除,因此,排除了亲本病毒污染的可能。该感染性克隆的构建,为深入探讨Asia1型口蹄疫病毒的致病机理以及新型疫苗的研制奠定了基础。

A full-length cDNA clone of foot-and-mouth disease virus （FMDV） Asial/YS/CHA/05 was constructed by assem- bling RT-PCR products in pBluescript II SK（＋） vector. The RNA transcripts was produced by in vitro transcription and transfected into BHK-21 cells. The rescued FMDV, which has a deleted Pst I site to differentiate contamination from parental FMDV, was successfully obtained. The availability of the cDNA clone would contribute to the understanding of molecular mechanisms of FMDV pathogenesis and the development of novel vaccines.