鸽新城疫病毒分离株F0裂解位点的基因克隆及序列分析

Molecular Cloning and Sequencing of the Cleavage Site of F0 Protein from the Isolate of Pigeon Newcastle Disease

为探索鸽新城疫的流行特点,在分子水平上阐明鸽新城疫病毒分离株与鸡新城疫毒株之间的相互关系,我们用RT-PCR技术获得了分离株FO裂解位点附近300bp的基因片段,将其克隆到pUC19载体上得到一重组克隆pNDV21,序列分析表明,插入片段与鸡新城疫强毒Texas相应区域的同源性为87.7%;而与弱毒株D26/76及La Sota的同源性分别是91.7%和98%.其FO裂解点的氨基酸顺序为Giy Arg-Gln-Gly-Arg.证明该分离株属于新城疫弱毒.

In order to investigate the epidemic characteristic of pigeon Newcastle disease and demonstrate the relation between isolates of pigeon and chicken Newcastle diseases, we got a 300 - base - pair fragment encoding the F2/F1 cleavage site of F0 protein from the pigeon - derived isolate by using RT- PCR technique. The fragment was cloned to pUC19 and one recombinant (pNDV21) was obtained. It was indicated by sequencing that the identity between the fragment and the same region of the virulent strain Texas was 87. 7 %, the identity with the avirulent strains D26/76 and La Sota were 91. 7% and 98% reapectively. The aminoacid sequence at the cleavage site of FO protein from the isolate was Gly - Arg - Gln ?Gly ?Arg. It could be concluded that the isolate wsa avirulent and close to La Sota.