Eukaryotic expression of enzymatically active human immunodeficiency virus type 1 reverse transcriptase

FEBS Letters

Kashuba, Vladimir I⁴  Pokholok, Dmitrii¹  Pokrovskaya, Katja⁴  Wahren, Britta²  Isaguliants, Maria G³  Kochetkov, Sergey N¹  Hinkula, Jorma² 

[1] W.A. Engelgardt Institute of Molecular Biology, Vavilova str 32, 117984 Moscow, Russia;Swedish Institute for Infectious Disease Control (SMI), Karolinska Institute, 171 82 Solna, Stockholm, Sweden;D.I. Ivanovsky Institute of Virology, Gamaleja str 16, 123098 Moscow, Russia;Molecular and Tumourbiology Center (MTC), Karolinska Institute, 17177 Stockholm, Sweden

关键词: Human immunodeficiency virus type 1;    Reverse transcriptase;    Eukaryotic expression;    DNA immunization;    IFL;    indirect immunofluorescent microscopy;    HIV-1;    human immunodeficiency virus type 1;    RT;    reverse transcriptase of HIV-1;    CMV IE;    cytomegalovirus immediate early promoter;   

DOI  :  10.1016/S0014-5793(99)00297-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Reverse transcriptase of human immunodeficiency virus type 1 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.

【 授权许可】

Unknown