Molecular cloning and expression in Escherichia coli of the glycoprotein gene of VHS virus, and immunization of rainbow trout with the recombinant protein

A recombinant viral haemorrhagic septicaemia virus glycoprotein expressed in insect cells induces protective immunity in rainbow trout

“…Concerning VHS, it was demonstrated very recently (Lorenzen et al, 1993) that the injection of rainbow trout with an almost complete VHSV G protein produced in E. coli could elicit specific anti-G antibodies. The recombinant protein had to be carefully renatured before injection.…”

Sequence variation of the glycoprotein gene identifies three distinct lineages within field isolates of viral haemorrhagic septicaemia virus, a fish rhabdovirus.

“…To perform such an analysis, we selected the G gene of VHSV as it codes for the only external viral protein, with the prospect that the genetic variability would then correlate with the serological variability, and thus help to resolve the serological classification of VHSV strains. Available sequence data on the VHSV G gene were limited to three strains from continental Europe (Thiry et al, 1991 ;Lorenzen et al, 1993), all belonging to serotype 1. To overcome this limitation, we determined the sequence of the G gene of 11 additional isolates of VHSV from continental Europe, the British Isles and North America.…”

Recombinant protein fragments from haemorrhagic septicaemia rhabdovirus stimulate trout leukocyte anamnestic responses in vitro

“…The RPE obtained in E. coli as inclusion bodies were difficult to handle and characterize (as also found by Lorenzen et al, 1993) because of their low yield and their minimal reaction with anti-VHSV protein MAbs (Table 1). This could be due to alterations in their conformation because of the formation of inclusion bodies and/or because of the presence of the fusion proteins needed for expression.…”

“…A total of six cysteine residues suggests that this region is highly important in the conformation of the protein. Since conformation and hydrophilicity, but not glycosylation, seem to be involved in the neutralizing epitope(s) of G (Lorenzen et al, 1993), it is possible that this region contains one of the neutralizing epitope(s).…”