山羊痘病毒反向末端重复序列的克隆与序列分析

应用PCR方法扩增山羊痘病毒QL、LD、Y、B-株反向末端重复序列片段,将其克隆至pMD18-T载体,构建了重组质粒pMD18-ITR,再将该重组质粒转化DH5α感受态细胞进行培养,对通过PCR、酶切鉴定为阳性的重组菌进行序列测定,并将所得序列与GenBank收录的2株山羊痘病毒、3株绵羊痘病毒全基因序列进行比较分析。结果:所测4个毒株序列与GenBank上收录的山羊痘病毒该片段核苷酸序列的同源性均为100%,与绵羊痘病毒该片段核苷酸序列的同源性均为98.3%。研究结果表明,山羊痘病毒反向末端重复序列与已收录的羊痘毒株之间该片段的同源性非常高,为今后兽医临床上应用PCR方法检测羊痘病毒提供了另一更为特异、保守的基因片段。

Clone and sequence analysis of inverted terminal repeat from goat poxvirus

The inverted terminal repeat gene fragments of goat poxvirus QL、LD、Y、B-strain were multiplied by PCR and cloned into the pMD18-T vector.The recombinant plasmid pMD18-ITR were constructed and then transformed into DH5α competent cells.The positive recombinant strains which were identified by PCR and restriction enzymes were sequenced,and compared with other complete genome sequences of two goat poxviruses and three sheep poxviruses available in the GenBank.The results indicated that the homology of nucleotide among four preserved viruses and two goat poxviruses were 100%,and 98.3% with three sheep poxviruses.The results above demonstrated that the homology was very high between the inverted terminal repeat gene fragment of goat poxvirus and capripoxvirus available in GenBank,and another more specific and conservative gene fragment was provided in detecting capripoxvirus by PCR in veterinary clinic in the future.