The invention relates to the technical field of biology, and particularly discloses a recombinant immunodeficiency plasmid and virus and application thereof. The immunodeficiency plasmid comprises LTR, gag gene, nucleotide sequence disclosed as SEQ ID No.1, vif gene, vpr gene, vpx gene, tat gene, rev gene, env gene and nef gene. The nucleotide sequence disclosed as SEQ ID NO.1 is introduced to the pathogenic SIVmac239 full-length plasmid frame to construct the immunodeficiency plasmid comprising a plurality of drug target spots, and packaging is performed in the 293T cell to generate immunodeficiency virus. The virus has the capacity for infecting target cells and the capacity of efficient replication, and is applicable to researching human immunodeficiency virus (HIV) in-vivo infection process, pathogeny characteristic, pathogenesis and immunoreaction and screening anti-HIV drugs and anti-HIV drug application strategies.