[Cloning, expression and preparation of polyclonal antibody for IBDV non-structure protein gene].
Infectious bursal disease virus (IBDV) VP5 gene was amplified and cloned into an N-terminal GST-Tag fusion expression vector, pGEX-4T-2, which was controlled by T7 promoter. The sequencing result showed that the VP5 gene was composed of 438 base pairs, and coded 145 amino acids. High VP5 product was expressed in E. coli BL21 induced by IPTG, and the GST-VP5 fusion protein existed in inclusion. High titer anti-VP5 serum was also prepared in New Zealand rabbit immunized with purified fusion protein inclusion. These results gave a basis for further research for VP5 function in IBDV replication and pathogenicity, which also paved the way for developing VP5 gene deleted IBDV live vaccine.
Yu
Lian
L
Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, China. yulian@zju.edu.cn
Li
Long
L
Liu
Yan
Y
Zheng
Jiang-tao
JT
Wei
Yong-wei
YW
Xu
Jian
J
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D007243:Infectious bursal disease virus / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; C518635:VP5 protein, infectious bursal disease virus; D017361:Viral Nonstructural Proteins