CLONING AND EXPRESSION OF RECOMBINANT HTLV-II ENVELOPE PROTEIN IN E. COLI

HUMAN T-LYMPHOTROPIC VIRUS TYPE II (HTLV-II) IS A HUMAN RETROVIRUS RELATED TO, BUT DISTINCT FROM, OTHER RETROVIRUSESTHAT ARE IDENTIFIED CAUSATIVE AGENTS OF DISEASE. LITTLE IS KNOWN ABOUT HTLV-II PREVALENCE IN THE POPULATION OR ITS DISEASE ASSOCIATIONS BECAUSE THE STUDY OF HTLV-II HAS LAGGED, IN PART DUE TO THE LACK OF READILY AVAILABLE, REASONABLY PRICED ASSAY REAGENTS. THIS PROJECT WILL CONSTRUCT RECOMBINANT CLONES OF THE HTLV-II PROTEINS IN AN EXPRESSION VECTOR BY POLYMERASE CHAINREACTION AMPLIFICATION AND ASSESS THE UTILITY OF THE RECOMBINANT PRODUCT AS SEROLOGIC REAGENTS FOR LARGE-SCALE PRODUCTION AND PURIFICATION. PHASE I OBJECTIVES INCLUDE CONSTRUCTION OF TWO CLONES OF THE HTLV-II ENVELOPE (ENV) GENE IN AN E. COLI EXPRESSION VECTOR AND PRELIMINARY CHARACTERIZATION OF THE EXPRESSED VIRAL PRODUCTS. IN PHASE II, ENV WILL BE CLONED INTO ADDITIONAL EXPRESSION SYSTEMS, AND THE MERITS OF THE DIFFERENT VECTORS WITH REGARDTO VIRAL PROTEIN PRODUCTION, ANTIGENICITY, AND SOLUBILITY WILL BE COMPARED. OTHER PHASE II OBJECTIVES INCLUDE CLONING OF THE BALANCE OF THE HTLV-II ANTIGENS AND DEVELOPMENT OF A PRELIMINARY PURIFICATION STRATEGY. RECOMBINANT VIRAL PROTEINS PRODUCED IN QUANTITY HAVING GOOD SOLUBILITY AND SEROLOGIC REACTIVITY CHARACTERISTICS ARE GOODCANDIDATES FOR RESEARCH AND BLOOD SCREENING ASSAY REAGENTS.