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    • 9. 发明授权
    • Stable TatIII cell lines, TatIII gene products, and assay methods
    • US4981790A
    • 1991-01-01
    • US806263
    • 1985-12-06
    • William A. HaseltineCraig A. RosenJoseph G. SodroskiWei C. Goh
    • William A. HaseltineCraig A. RosenJoseph G. SodroskiWei C. Goh
    • C12P19/34C07K14/15C07K14/16C12N5/10C12N15/00C12N15/09C12N15/85C12P21/00C12R1/91
    • C07K14/005C12N15/85C12N2740/14022C12N2740/16322
    • This invention describes stable tat.sub.III cell lines. It is disclosed that by transfecting a preselected tat.sub.III cell line with a vector containing a sufficient amount of the HTLV-III LTR responsive to tat.sub.III gene products for trans-activation and an enhancer upstream of the tat.sub.III responsive segment, it is possible to express high levels of the tat.sub.III gene products. By including a preselected heterologous gene on this vector, it is also possible to express high levels of a desired gene product. A substantially pure protein comprising 86 amino acids and having an apparent molecular weight of about 14,000 dalton and exhibiting trans-activating activity is also disclosed. This protein and polypeptides having trans-activating ability, which is also disclosed, can be used to produce high levels of a desired gene product. A method of detecting the presence of HTLV-III/LAV virus in an individual is also disclosed and comprises the step of:(a) incubating whole blood or lymphocytes from the invididual to be tested with tat.sub.III cell lines of the present invention in a culture medium; and(b) screening for cytopathic effects on the cells is also disclosed. A method of screening for a compound that inhibits trans-activation of the tat.sub.III gene product is also disclosed and comprises the steps of:(1) transacting a tat.sub.III cell line of the present invention with a vector containing a gene that expresses a selectable marker and whose expression is under the control of an HTLV-III LTR;(2) transfecting the same type of tat.sub.III cell lines as in step (1) with the selectable gene chosen in step (1) but under the control of a different regulatory sequence;(3) thereafter, adding a preselected compound to each of the cell lines in increasing concentrations; and(4) measuring the expression of the selectable gene product to determine whether the compound effects the tat.sub.III function without being toxic to the cell.