Generation and evaluation of a recombinant myxomavirus expressing the VP60 protein of rabbit haemorrhagic disease virus / 生物工程学报

Rabbit haemorrhagic disease virus (RHDV) and myxoma virus (MYXV), are two pathogens that have harmful effect on rabbit breeding and population decline of European rabbits in their native range, causing rabbit haemorrhagic disease (rabbit fever) and myxomatosis, respectively. The capsid protein VP60 of the RHDV represents the major antigenic protein. To develop a recombinant bivalent vaccine candidate that can simultaneously prevent these two diseases, we used the nonessential gene TK (thymidine kinase) of MYXV as the insertion site to construct a recombinant shuttle vector p7.5-VP60-GFP expressing the RHDV major capsid protein (VP60) and the selectable marker GFP. Then the shuttle vector p7.5-VP60-GFP was transfected into rabbit kidney cell line RK13 which was previously infected with MYXV. After homologous recombination, the recombinant virus expressing GFP was screened under a fluorescence microscope and named as rMV-VP60-GFP. Finally, the specific gene-knock in and expression verification of the vp60 and gfp genes of the recombinant virus was confirmed by PCR and Western blotting. The results showed that these two genes were readily knocked into the MYXV genome and also successfully expressed, indicating that the recombinant MYXV expressing the vp60 of RHDV was generated. Protection against MYXV challenge showed that the recombinant virus induced detectable antibodies against MYXV which would shed light on development of the effective vaccine.

Advances in molecular biology research on human parvovirus B19 / 生物工程学报

Human parvovirus B19 (B19 virus) is one of the two parvoviruses that cause human diseases. As an important pathogen to humans, it causes infectious erythema in children, acute aplastic anemia, fetal edema and death. In this review, we focus on the recent advances in the molecular virology of B19V, such as viral genotypes, viral receptor, genomic features and viral replication, viral transcription and post-transcription regulation, viral nonstructural and structural protein features and functions, viral diagnosis and antiviral agents, to provide reference for further study of B19 pathogenesis mechanisms, treatment and diagnostic strategies.

Regulatory effect of 11 kDa protein of parvovirus B19 on NF-kappaB pathway in Hela cells / 生物工程学报

The 11 kDa protein, a small nonstructural protein of parvovirus B19, may play important roles in viral replication cycle. To investigate the effect of 11 kDa protein on the NF-kappaB signaling pathway, we first prepared the poly-antiserum using GST-11 kDa fusion protein purified via prokaryotic expression system, and demonstrated that the 11 kDa protein mainly localized in cytoplasm when expressed in Hela cells. Meanwhile, luciferase activity assay and Western blotting assay showed that 11 kDa up-regulated the transcriptional activity of NF-kappaB and induced the degradation of IkappaB-alpha in Hela cells. Moreover, the 11 kDa protein activated the IL6 promoter, which is probably through the NF-kappaB pathway. Taken together, these results suggested that 11 kDa protein may contribute to activating inflammatory factors through participating in the cell signaling pathway.