Objective To analyze 9 nucleoprotein gene sequences of rabies viruses circulating in Henan province, China, and to explore the probable mutant on their pathogenicity and virulence. Methods N gene sequences of 9 rabies viruses derived from Henan province were amplified by RT-nested-PCR, and then were cloned and sequenced. Bioinformatics software was used to determine the genetic characteristics of these rabies viruses. Results The sequences of N gene of nine Henan isolates were identical, with homogeny between 97.5%-99.3% and 98.4%-99.8% on nucleotide and deduced amino acid level, respectively. They were much more homogeneous with CTN than with other vaccine strains, with the identities being 88.9%-90.1% and 97.6%-98.0% on nucleotide and amino acid, respectively, while the identities between them and other vaccine strains were 84.4%-87.9% and 94.2%-97.3%. There were several amino acid substitutions in the nucleoprotein of 9 Henan isolates, compared with the known genotype 1 isolates. Conclusions The rabies viruses derived from Henan province were still genotype 1. These isolates of rabies virus were diverged from the vaccines in both gene sequences and deduced amino acid sequences, and these divergences might result in the inefficiency of the vaccines.