Thereversetranscriptase(RT)proteinofhepatitisBvirus(HBV)hasbeensuccessfullyexpressedbyrecombinanttechnologyinEschericahiacoli(E.coli).Inthisstudyweaimedtodevelopasemi-quantitativeassayforthestudyofHBVRTproteinusingthissystem.CompleteHBVpolymerasegenefromawildtypevirus(rt306P)andthepolymerasegenefromamutant,withrt306Psubstitutedbyserine(rtP306S)wereseparatelyfusedtothemaltosebindingprotein(MBP)geneandexpressedinE.colirespectively.TheexpressionlevelsofHBVpolymerasegenesfromthewildtypevirusanditscounterpartmutantatrt306werecompared.Whentheseproteinsweresemi-quantifiedbyWesternblottingusingrabbitanti-TPserum,thertP306SmutantshoweddecreasedexpressionofMBP-HBVpolymerase.Bythismethod,wehaveshownthattheexpressionlevelofHBVRTcouldbeaffectedbysubstitutionsinitsaminoacidsequences,andthismethodcouldbeusedtostudythecharacteristicsofHBVRTprotein.
