[Study on the self-assembly ability of expressed capsid protein of rabbit haemorrhagic disease virus that fused with foreign epitope at the N-terminus].
The RHDV capsid protein (VP60) gene was first subcloned into the transfer plasmid pBLUEBACHIS2B located downstream of a 6 * HIS tag, then the recombinant transfer plasmid DNA were cotransfected Sf 9 cells with Bac-N-Blue DNA and purified for cloned recombinant baculovirus by plaque assay. The expression of fused-VP60 gene was analyzed by SDS-PAGE and Western blot. A specific 69 kD protein band was obtained. Observed under electron micrography, the recombinant baculovirus expressed VP60 protein assembled into viruslike particles which were morphologically and antigenically similar to native RHD virus but did not package RNA. The close-to-native conformation of the VLPs was also supported by the haemagglutination test, in which recombinant VLPs, like RHDV, agglutinated human blood type O erythrocytes.
Yan
Wei-wei
WW
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. yzuyww@yahoo.com
Cui
Zhi-zhong
ZZ
Wang
Yong-kun
YK
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D000939:Epitopes / Q000276:immunology; D017929:Hemorrhagic Disease Virus, Rabbit / Q000235:genetics* / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D018411:Spodoptera; D015678:Viral Structural Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D019065:Virus Assembly
D000939:Epitopes; D011994:Recombinant Proteins; D015678:Viral Structural Proteins; C081149:viral protein 60, rabbit hemorrhagic disease virus