Michitaka Yamamoto

Construction of recombinant Marek's disease virus type 1 (MDV1) expressing the Escherichia coli lacZ gene as a possible live vaccine vector: the US10 gene of MDV1 as a stable insertion site

This paper describes the construction of a recombinant Mareks disease virus serotype 1 (MDV1) in which the Escherichia coli lacZ gene was inserted into the open reading frame homologous to the US10 gene of herpes simplex virus 1 (HSV1). The recombinant virus replicated as well in cell culture as the parental MDV1 K-554 strain. Chickens immunized with the virus were protected against challenge with virulent MDV1, and produced a high level of antibodies against beta-galactosidase as well as against MDV1 antigens. The antibody titres persisted for at least 16 weeks. These results demonstrate that the US10 gene of MDV1 is an effective site for the insertion of foreign genes from which to construct a polyvalent live vaccine for poultry.

Studies on the antigenicity and nucleotide sequence of the rabies virus Nishigahara strain, a current seed strain used for dog vaccine production in Japan.

The Nishigahara strain of rabies virus, a current seed strain used for animal vaccine production in Japan, is believed to derive from the original Pasteur strain obtained from Paris in or before 1915. In Japan, the virus was serially passaged through several kinds of animals and cell cultures. Reactions with anti-nucleocapsid protein monoclonal antibodies (MAb-N) indicated the Nishigahara strain had maintained the antigenic profile of the Pasteur virus. Reactions with monoclonal antibodies to the glycoprotein (MAb-G) revealed differences between the Nishigahara strain and the Pasteur strain; however, the Nishigahara strain maintained a closer resemblance to the Pasteur virus than to other Pasteur-related viruses or to rabies strains unrelated to the Pasteur strain. Comparative amino acid sequence analysis of cloned cDNA encoding the G gene confirmed the antigenic differences among these strains and the resemblance of the Nishigahara strain to the original Pasteur strain. Comparative nucleotide sequence analysis of the noncoding pseudogene region (Tordo et al., Proc Natl Acad Sci USA83, 3914–3918, 1986) revealed different relationships. Unlike the Pasteur strain, which encodes a transcription-terminating signal at the end of the G gene (marking the beginning of the pseudogene), a long G-L intergenic sequence in the Nishigahara strain was connected to the 3′ end of the cDNA, and the transcription-terminating signal was present only at the end of, but not before, the pseudogene. These results are not inconsistent with the documented origin of the Nishigahara strain, but the genome structure around the pseudogene region suggests divergence from the Pasteur strain and a closer resemblance to other strains of rabies virus.