Preparation of ChIL-2 and IBDV VP2 fusion protein by baculovirus expression system.
This study aims to produce an effective subunit vaccine against infectious bursal disease virus (IBDV). The genes of chicken interleukin-2 (ChIL-2) and IBDV viral protein 2 (VP2) were amplified and fused by splice overlap extension-polymerase chain reaction (SOE-PCR). The fusion gene was digested by EcoR I/Kpn I and inserted into pBacPAK8 vector, resulting in recombinant transfer plasmid pBacPakVP2-IL2. The recombinant plasmid was transfected into Sf-9 cells accompanied with hybrid nuclear polyhedrosis virus (HyNPV) genome DNA and lipofectin. Plaque-purification indicated that we had got the recombinant Hy-VP2-IL2. Fusion protein VP2-IL2 was expressed effectively both in insect cells and bombyx mori. The expression of fusion protein was confirmed by ELISA, SDS-PAGE and Western blotting assay, respectively. This efficient system allows us to meet the need for inexpensive vaccines required by the poultry industry.
Liu
Yan
Y
Department of Biomedical Engineering, Zhejiang University, Hangzhou, 310027, China.
Wei
Yongwei
Y
Wu
Xiaofeng
X
Yu
Lian
L
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology; D015153:Blotting, Western; D012831:Bombyx / Q000166:cytology / Q000821:virology; D004274:DNA, Recombinant / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D015678:Viral Structural Proteins / Q000096:biosynthesis* / Q000235:genetics*
D004274:DNA, Recombinant; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; C082513:VP2 protein, infectious bursal disease virus; D015678:Viral Structural Proteins