[The expression of IBDV VP2 in chicken primary myoblast cells using the baculovirus vector].
OBJECTIVE
To construct the recombinant baculovirus expressing Infectious bursal disease (IBDV) VP2 gene in the chicken primary myoblast cells.


METHODS
A proteinase K digestion and phenol-chloroform extraction method was used to extract dsRNA genome from IBDV. VP2 gene was amplified by Reverse Transcription Polymerase Chain Reaction (RT-PCR) with the genome RNA as template. The pFastBac-pCMV-VP2 baculovirus transfer vector was constructed by inserting VP2 gene under the immediate-early promoter of cytomegalovirus. The VP2 recombinant bacmid was obtained by Bac-to-Bac system and transfected sf9 insect cell to acquire VP2 recombinant baculovirus. After amplification of recombinant baculovirus on cell passages, the recombinant virus was seeded on chicken primary myoblast cells with 50 multiplicity of infection (MOI), and the cells were harvested at 72 hours after infection.


RESULTS
Sodium Dodecyl Sulphate Poly-Acrylamide Gel Electrophoresis (SDS-PAGE) and Western blot results showed that the VP2 gene was successfully expressed in chicken primary myoblast cells. The product was a 48kDa protein and could be recognized by anti-IBDV serum.


CONCLUSIONS
The recombinant baculovirus could efficiently delivery IBDV VP2 gene into chicken primary cells and that CMV, a mammalian-cell-active promoter, was functional in chicken primary cells and could direct the expression of VP2 antigen protein. The research can be a potential basis for the development of baculovirus vector vaccines for IBDV and other avian infectious disease.
Ge
Jingping
J
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, China. gejingping@126.com
Gao
Dongni
D
Lou
Zhuangwei
Z
Ping
Wenxiang
W
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D002645:Chickens; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000502:physiology; D007243:Infectious bursal disease virus / Q000235:genetics / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism* / Q000821:virology; D018411:Spodoptera; D015678:Viral Structural Proteins / Q000235:genetics* / Q000378:metabolism
C082513:VP2 protein, infectious bursal disease virus; D015678:Viral Structural Proteins