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Virologica Sinica, 18 (2) : 159-163, 2003
余晓岚, 肖少波,方六荣,金梅林,陈焕春**
Animal Science and Veterinary Medicine College,Huazhong Agricultural University,Wuhan,430070,China
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.
  Published online: 15 Apr 2003
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