鸭乙型肝炎病毒复制复合体（DHBV RCs）的提纯及其对外源模板的利用

Isolation,Purification and DNA Polymerase Activity Analysis of DHBV Replicative Complexes

鸭乙型肝炎病毒复制复合体（ＤＨＢＶＲＣｓ）的提纯及其对外源模板的利用邵兴无，陶佩珍，郭巨涛，陈鸿珊（中国医学科学院医药生物技术研究所，北京１０００５０）关键词鸭乙型肝炎病毒复制复合体，ＤＮＡ聚合酶，核心抗原，外源模板１鸭乙型肝炎病毒复制复合体（ＤＨＢ...

uck hepotitis B virus repl icative cumplexes(DHBV RCs)were isolated from DHBV-infectedduck liver by clironlatography through Bio-Gel A 5m followed by sucrose gradient centrifugation.Therelative endogenous DNA polynierase activity of the purified RCs was l00 times higlier than that of theunchronlatographed one, confirined by detecting their DHBcAg with Dot-EIA method.The utilization of exogenous template in reverse transe,riptale reaction was determined by incurporation of ￣3H dTTP into poly(rA)(dT)_(12-18).The resul ts indicated that DHBV RCs could use the exog.nous template,the activity was about 40% of that of the endugenous DNA polymerase,Pretreatmentof DHBV RCs with nonionic detergent(5% Triton X-100),high concentration saIt(1 mol/L NaCl)and low pH(0.06mol/L Tris/glycine pH2.5)did not enhance the utilimtion of exogenous templateactivity.