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

Rina Rosin-Arbesfeldb, Pnina Mashiah, Dieter Willbold, Paul Rosch, Steven R. Tronick, Abraham Yaniv, Arnona Gazit*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

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.

Original languageEnglish
Pages (from-to)307-311
Number of pages5
JournalGene
Volume150
Issue number2
DOIs
StatePublished - 1994

Keywords

  • EIAV
  • His tag
  • Recombinant protein
  • frans-activation
  • lentiviruses
  • pET prokaryotic expression vector

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