Hepadnavirus polymerase gene product having RNA-dependent DNA pr

Details Hepadnavirus polymerase gene product having RNA-dependent DNA pr Hepadnavirus polymerase gene product having RNA-dependent DNA pr

1992-08-28

1994-08-02

Wax, Robert A.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

435 6, 435 15, 435 691, 435 9151, 935 14, 935 17, 25218311, 536 232, 536 2372, C12N 912, C12N 1510, C08L 500, C12Q 148

Patent

active

053345258

ABSTRACT:
An enzymatically active, substantially virus-free hepadnavirus polymerase gene product is provided, which possesses a DNA priming function and reverse transcriptase activity, both of which are dependent on the presence of an RNA template and magnesium ions. The hepadnavirus polymerase gene product is produced by in vitro transcription and translation of a hepadnavirus pol gene.
The enzymatically active hepadnavirus polymerase gene product is used in rapid in vitro assays to screen potential anti-hepadnaviral agents. An assay for the DNA priming activity of the gene product is provided, wherein covalent attachment of labelled dGTP to the gene product is quantitatively measured. An assay for the reverse transcriptase activity of the gene product is provided, wherein incorporation of nucleotide triphosphates into the nascent DNA minus strand is measured, taking advantage of the covalent attachment of said nascent strand to the polymerase gene product.

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