余晓岚， 肖少波，方六荣，金梅林，陈焕春**

A 0．77kb UL49 gene encoding the tegument protein VP22 was amplified from cells culture infected with Bovine herpesvirus 1 (BHV—I)bv PCR．The DNA fragment was further cloned into pM D 1 8一T vector,resulting in recombinant plasmid pM D—VP22．The sequences were determined an d there is no diference with the sequences of the 【，L 9 gene of BHV—I Cooper strain ．Th e 0．77kb fragment was released from plasmid pMD．．VP22 an d subcloned into the vectors pET．．28a an d pEGFP-C 1 respectively，to genera~ a prokaryotic expression pET一28aVP22 an d eukaryotic expression plasmid pEGFPVP22．pET一28aVP22 was consquenfly transformed into E．coli BL21(DE3)．After induced with IPTG．the fusion protein was expressed an d the molecular weight was about 38l(Da． pEGFPVP22 Was tran sfected into PK一15 cells an d the clone cells were selected out wim G418．Th e autofluorescence of the clone cells line tran sfected with pEGFPVP22 could be observed under inve~ed fluorescence microscope．It is interesting that the fluorescence primarily targeted the nucleus of cells whereas the autofluorescence of cells tran sfected with control vector pEGFP—C1 localized throughout the cytoplasm．