Objective To explore the effects of M gene rearrangement of rabies virus HEP-Flury on gene transcription and protein expression, reveal the correlation between phenotypic consequences of virus in mouse neuroblastoma (NA) cells and M gene rearrangement.Method The gene transcription, expression, growth and spread of parent strain rHEP-Flury and M gene rearranged virus stains (M2 and M4) in infected NA cells were compared by fluorescence quantitative PCR, Western blot, growth and spread experiments of virus in NA cells.Result The transcription and expression of rabies virus structural gene were mainly affected by RNA synthesis of virus genome. The gene position mainly affected the ratio of transcription in a complete transcription process. The transcription ratios of leader RNA (LeRNA) and L mRNA of M gene rearranged viruses were significanthy higher than those of rHEP-Flury. The growth and spread of M gene rearranged viruses in NA cells were inferiorer than rHEP-Flury.Conclusion rHEP-Flury has wild-type gene order of rabies virus. Its growth and spread in NA cells are superiorer to M gene rearranged viruses. The position of structural gene in genome determines its transcription ratio in a complete transcription process and affects the growth and spread of virus in NA cells.