Patent ID: 6593082
Filing Date: 2003-07-15
Classification: C12N,C12Q

Abstract:
A method for assaying human hepatitis B virus (HBV) DNA polymerase in vitro, comprising:providing a 5â€² DNA oligonucleotide comprising the SP6 viral polymerase promoter and an oligonucleotide sequence matching the 5â€² end of the human HBV DNA polymerase coding region; providing a 3â€² oligonucleotide comprising an oligonucleotide sequence matching the 3â€² end of the human HBV DNA polymerase coding or non-coding region; contacting the 5â€² oligonucleotide and 3â€² oligonucleotide with a sample containing a DNA encoding human HBV DNA polymerase; performing PCR amplification under conditions that allow amplification of the DNA encoding human HBV DNA polymerase to produce PCR products; transcribing and translating the PCR products in eukaryotic cell-free lysates to produce in vitro translated human HBV DNA polymerase; and measuring the priming activity of the in vitro translated HBV DNA polymerase in the presence of an agent labeled with a detectable marker by detecting the amount of the agent incorporated into a complex formed between the in vitro translated human HBV DNA polymerase and the agent.