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    • 3. 发明申请
    • RECOMBINANT EXPRESSION OF hHBV REVERSE TRANSCRIPTASE (RT)
    • hHBV逆转录酶(RT)的重组表达
    • WO2002088343A2
    • 2002-11-07
    • PCT/EP2002/004642
    • 2002-04-26
    • PHYTRIX AGBUCHNER, JohannesMUSCHLER, PaulHALSBECK, Martin
    • BUCHNER, JohannesMUSCHLER, PaulHALSBECK, Martin
    • C12N9/12
    • C12N9/1276C07K2319/00
    • The present invention relates to a method of producing a functional cell-free hepatitis B virus (HBV) reverse transcriptase (RT) comprising the steps of expressing HBV-RT in E. coli cells employing a suitable expression plasmid, or in a cell-free transcription-translation system and if expression was carried out in E. coli cells, lysing said E. coli cells and purifying HBV-RT from E. coli lysate or from said transcription-translation system. The invention further relates to a method of screening for an inhibitor of HBV-RT activity comprising the steps of contacting the cell-free HBV-RT produced according to methods of the present invention and with a potential inhibitor and assaying whether said potential inhibitor inhibits HBV-RT activity. Furthermore, the invention provides for a method of producing a pharmaceutical composition comprising the step of formulating the inhibitor identified by the screening method of the invention into a pharmaceutical composition.
    • 本发明涉及一种生产功能性无细胞乙型肝炎病毒(HBV)逆转录酶(RT)的方法,包括以下步骤:使用合适的表达质粒在大肠杆菌细胞中表达HBV-RT,或以无细胞 转录翻译系统,并且如果在大肠杆菌细胞中进行表达,则将大肠杆菌细胞裂解并从大肠杆菌裂解物或所述转录翻译系统中纯化HBV-RT。 本发明还涉及一种筛选HBV-RT活性抑制剂的方法,包括以下步骤:将根据本发明方法产生的无细胞HBV-RT与潜在的抑制剂接触,并测定所述潜在抑制剂是否抑制HBV -RT活动。 此外,本发明提供了制备药物组合物的方法,其包括将通过本发明的筛选方法鉴定的抑制剂配制成药物组合物的步骤。