title: Eukaryotic expression of enzymatically active human immunodeficiency virus type 1 reverse transcriptase.
abstract: Reverse transcriptase of human immunodeficiency virus type I is a vitalenzyme in the HIV-1 replication cycle and an attractive target of attempts to arrest a primary viral infection. We designed a vector for eukaryotic expression of the 66 kDa subunit of reverse transcriptase under the control of the immediate early cytomegalovirus promoter. Efficient transient expression of the 66 kDa subunit of reverse transcriptase was achieved in a variety of cells. Immunostaining of the transfected cells revealed the cytoplasmatic localization of reverse transcriptase. Reverse transcriptase activity was detected in all transfected cell lines. Injection of this plasmid encoding the 66 kDa subunit of reverse transcriptase into mice resulted in strong reverse transcriptase-specific immune responses indicating that the 66 kDa subunit of reverse transcriptase is expressed in vivo. Sera from DNA-immunized mice inhibited reverse transcription in vitro.
authors.affiliation: D.I. Ivanovsky Institute of Virology, Moscow, Russia. maria.isaguliants@solna.mail.telia.com
authors.forename: M G
authors.initials: MG
authors.lastname: Isaguliants
authors.forename: K
authors.initials: K
authors.lastname: Pokrovskaya
authors.forename: V I
authors.initials: VI
authors.lastname: Kashuba
authors.forename: D
authors.initials: D
authors.lastname: Pokholok
authors.forename: J
authors.initials: J
authors.lastname: Hinkula
authors.forename: B
authors.initials: B
authors.lastname: Wahren
authors.forename: S N
authors.initials: SN
authors.lastname: Kochetkov
mesh_terms: D016475:3T3 Cells; D016915:AIDS Vaccines / Q000235:genetics / Q000276:immunology; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D005822:Genetic Vectors; D015483:HIV Antibodies / Q000097:blood; D054303:HIV Reverse Transcriptase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D007114:Immunization; D051379:Mice; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D011487:Protein Conformation; D014162:Transfection
chemical_list: D016915:AIDS Vaccines; D004274:DNA, Recombinant; D015483:HIV Antibodies; D054303:HIV Reverse Transcriptase