运用锚引物扩增新城疫病毒鹅源分离株基因组末端序列

AMPLIFICATION OF THE GENOMIC TERMINUS OF NEWCASTLE DISEASE VIRUS OF GOOSE ORIGIN BY ANCHOR PRIMERS

在T4RNA连接酶的作用下,将人工合成的锚引物直接与新城疫病毒(NDV)鹅源毒株ZJ1株的基因组5′末端的反转录产物及NDV基因组3′末端直接相连,然后用聚合酶链反应(PCR)和RT PCR分别快速扩增基因组5′末端和3′末端。测序结果表明:3′末端的Leader序列为55nt,5′末端的trailer序列为114nt。将该序列与GenBank报道的NDV鸡源毒株的相应序列相比,ZJ1株与6个鸡源NDV毒株的Leader和trailer序列的同源性分别为83.6%～92.7%和60 5%～63.2%。序列比较结果表明:该鹅源毒株与鸡源毒株在基因组3′端调控区有较高的同源性,而在基因组5′端的调控区同源性则较低。

The 3′terminus and the reverse transcription product of the 5′terminus of the genome of Newcastle disease Virus (NDV) of goose origin (ZJ1) were ligated with an artificial synthesized anchor primer by T4 RNA ligase, then RTPCR and PCR were utilized to rapidly amplify the 3′terminus and 5′terminus of NDV genome. The sequencing results of the PCR products showed that the leader and trailer sequence of ZJ1 were 56 and 114nt long respectively. Then DNASTAR program was utilized to compare the 3′terminus and 5′terminus of ZJ1 with those of chicken origin released in GenBank, the results showed the homology of the leader of ZJ1 and 6 NDV of chicken origin ranged in 83.6%～92.7%, the homology of the trailer sequence of ZJ1 and 6 NDV of chicken origin ranged in 60.5%～63.2%. The comparing result showed that there was high homology in the leader sequence between ZJ1 and NDV of chicken origin; while the homology of trailer sequence of ZJ1 with those of chicken origin was low.