Expression of the Capsid Precursor Protein gene of Foot-and-mouth Disease Virus and Green Fluorescent Protein Gene in BHK-21 Cells Mediated by Retroviral Vector

We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.

Microcalorimetric Study of Energy Metabolism of HSV-2 and FMDV Infection Processes

The metabolic thermogenic power data of the HSV-2 infected HeLa cells and the FMDV infected BHK-21 cells were determined by LKB-2277 bioactivity monitor. The aim of the study was to investigate the difference of the cell metabolism under the action of two different viruses and the effects of hyperthermia and drugs on it. The results illustrated that the metabolic thermogenic power of infected cells was larger than the uninfected ones and there was a significant difference between the metabolism heat released by the two types of infected cells. From the maximal thermal power and total metabolism heat, the infection process was observed to be thermosensitive and could be inhibited by interferon. Our experiments also revealed that 6 month storage of FMDV could attenuate its virulence and infectivity. The study shows that microcalorimetry is a potent tool to investigate the metabolism of virus infection process.