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Biological activity and intracellular location of the Tat protein of equine infectious anemia virus |
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| Authors: | Rina Rosin-Arbesfeldb Pnina Mashiah Dieter Willbold Paul Rosch Steven R Tronick Abraham Yaniv Arnona Gazit |
| Institution: | a Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Tel Aviv 69978, Israel b Lehrstuhlfur Struktur und Chemie der Biopolymere, Universität Bayreuth, D-95440, Bayreuth, Germany. Tel. (49-921) 55-3540 c Laboratory of Cellular and Molecular Biology, National Cancer Institute, Bethesda, MD, 20892, USA. Tel. (1-301) 496-9683 |
| 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. |
| Keywords: | Recombinant protein EIAV lentiviruses frans-activation His tag pET prokaryotic expression vector |
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