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The regulatory and accessory proteins of human immunodeficiency virus type 1 (HIV-1) are produced from singly or multiply spliced mRNAs. We have used HIV-1-specific oligonucleotide primer pairs in a polymerase chain reaction procedure on RNA from lymphocyte cell lines infected with HIV-1(228,200). The amplified products were analysed by hybridization with splice junction-specific oligonucleotide probes to determine splice donor/splice acceptor combination utilization, subcloned into a plasmid vector and the nucleotide sequences were obtained. Two novel splice acceptor sites have been identified.