Elsevier

Gene

Volume 150, Issue 2, 1994, Pages 307-311
Gene

Biological activity and intracellular location of the Tat protein of equine infectious anemia virus

https://doi.org/10.1016/0378-1119(94)90443-XGet rights and content

Abstract

The Tat protein of equine infectious anemia virus (EIAV) was synthesized in Escherichia coli using the inducible expression plasmid, pET16b, which contains a His.Tag leader, thus allowing for rapid and efficient enrichment of the histidine-tagged protein by metal affinity chromatography. Yields of up to 20 mg of Tat were obtained from 1011 bacterial cells. The recombinant Tat protein was shown to potently trans-activate the EIAV long terminal repeat (LTR) following its introduction into canine cells by ‘scrape loading’. The EIAV Tat protein was found to localize predominantly within the cytoplasm, in contrast to HIV-1 Tat. The availability of large amounts of purified functional EIAV Tat protein should greatly facilitate detailed structure-function analyses.

References (32)

  • D. Derse et al.

    A minimal lentivirus Tat

    J. Virol.

    (1991)
  • L. Dorn et al.

    Cis- and trans-acting regulation of gene expression of equine infectious anemia virus

    J. Virol.

    (1988)
  • L. Dorn et al.

    Equine infectious anemia virus tat: Insights into the structure, function and evolution of lentivirus trans-activator proteins

    J. Virol.

    (1990)
  • A.D. Frankel et al.

    Tat protein from immunodeficiency virus forms a metal-linked dimer

    Science

    (1988)
  • R. Gentz et al.

    Bioassay for trans-activation using purified human immunodeficiency virus tat-encoded protein; trans-activation requires mRNA synthesis

  • M.A. Graeble et al.

    Human immunodeficiency virus type 1 transactivator protein, Tat, stimulates transcription read-through of distal terminator sequences in vitro

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