prokaryotic expression, purification, and polyclonal antibody production of a truncated recombinant rabies virus l protein

background: rabies virus (rabv) is a deadly neurotropic virus that causes the disease of rabies in humans and animals. l protein is one of the large structural protein of rabies virus, which displays multiple enzymatic activities, and is required for viral transcription and replication.objectives: a truncated l protein of rabies virus is being cloned, expressed and purified to produce relevant polyclonal antibody.materials and methods: the gene fragment of l protein of rabv was subcloned into prokaryotic expression vector pbackground: rabies virus (rabv) is a deadly neurotropic virus that causes the disease of rabies in humans and animals. l protein is one of the large structural protein of rabies virus, which displays multiple enzymatic activities, and is required for viral transcription and replication.objectives: a truncated l protein of rabies virus is being cloned, expressed and purified to produce relevant polyclonal antibody.materials and methods: the gene fragment of l protein of rabv was subcloned into prokaryotic expression vector pet-28a and transformed into e. coli rosetta de3 host strain. the recombinant l protein of rabv was expressed and characterized by sds-page and western blot analysis using anti-his tag antibody. mice were immunized with the purified recombinant l protein, the reaction of the anti-serum was checked by immunofluorescence and dot-blot, respectively.results: the results of pcr and sequencing confirmed that the fragment of l gene of rabv was successfully cloned into the expression vector. the expression of recombinant l protein fragment induced by iptg was confirmed by the band of 43 kda in sds-page and western blot. the antiserum of purified l protein immunized mice was reacted with rabv infected n2a cells and suckling mouse brain tissue lysates. conclusions: our data showed that the recombinant l protein produced by pet-28a vector was very successful, and the purified l protein could efficiently induce the antibody response in mice. the antiserum could recognize the virus in rabv infected cells and tissue very well.et-28a and transformed into e.coli rosetta de3 host strain. the recombinant l protein of rabv was expressed and characterized by sds-page and western blot analysis using anti-his tag antibody. mice were immunized with the purified recombinant l protein, the reaction of the anti-serum was checked by immunofluorescence and dot-blot, respectively.results: the results of pcr and sequencing confirmed that the fragment of l gene of rabv was successfully cloned into the expression vector. the expression of recombinant l protein fragment induced by iptg was confirmed by the band of 43 kda in sds-page and western blot. the antiserum of purified l protein immunized mice was reacted with rabv infected n2a cells and suckling mouse brain tissue lysates. conclusions: our data showed that the recombinant l protein produced by pet-28a vector was very successful, and the purified l protein could efficiently induce the antibody response in mice. the antiserum could recognize the virus in rabv infected cells and tissue very well.