PMID: 7539509Jul 1, 1995Paper

Nucleotide priming and reverse transcriptase activity of hepatitis B virus polymerase expressed in insect cells

Journal of Virology
R E LanfordB Beames

Abstract

Hepadnavirus polymerases initiate reverse transcription in a protein-primed reaction that involves the covalent linkage of the first deoxyribonucleotide to the polymerase polypeptide. Analysis of the initial steps in this reaction as well as certain details of genome replication has been hampered by the difficulties encountered in the expression of functional hepadnavirus polymerases in heterologous systems. We have expressed human hepatitis B virus (HBV) polymerase (pol) in insect cells, using the recombinant baculovirus system. Analysis of immunoaffinity-purified pol indicated that (i) a portion of pol had initiated minus-strand DNA synthesis within infected insect cells; (ii) the pol mRNA appeared to be the template for reverse transcription; (iii) the products were small (100 to 500 nucleotides); (iv) only minus-strand DNA was synthesized; (v) the products were covalently bound to protein; and (vi) the 5' end of the minus-strand DNA mapped to DR1 by primer extension. The purified pol was also active in an in vitro polymerase assay. Analyses suggested that a different fraction of pol was active in the in vitro assays. Incubation of pol with labeled deoxyribonucleotide triphosphates resulted in the labeling of the pol polypep...Continue Reading

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Citations

Mar 8, 2000·Microbiology and Molecular Biology Reviews : MMBR·C Seeger, W S Mason
Feb 7, 2002·Expert Opinion on Investigational Drugs·William DelaneyStephen A Locarnini
May 11, 2002·Journal of Cellular and Molecular Medicine·C Kreutz
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Jan 28, 2005·The Journal of Alternative and Complementary Medicine : Research on Paradigm, Practice, and Policy·James R JacobYoung-Ho Kim
Jul 21, 2007·Journal of Huazhong University of Science and Technology. Medical Sciences = Hua Zhong Ke Ji Da Xue Xue Bao. Yi Xue Ying De Wen Ban = Huazhong Keji Daxue Xuebao. Yixue Yingdewen Ban·Xiaoyan WangJusheng Lin
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Apr 17, 1997·Biochemical and Biophysical Research Communications·Y I LeeG Jung

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