Expression of the active human and duck hepatitis B virus polymerases in heterologous system of Pichia methanolica.
We expressed the Hepatitis B virus polymerase (HBV P protein) using a recently introduced yeast system, Pichia methanolica. HBV (1-680 amino acids) and Duck Hepatitis B virus (DHBV, 1-780 amino acids) polymerase were expressed and showed DNA dependent DNA polymerase (DDDP). The DHBV polymerase had RNA dependent DNA polymerase (RDDP) and RNase H activities. We present a new simplified way of obtaining active viral P protein using the yeast expression system. The viral P proteins proved to be stable and were not aggregated in the yeast system.
Choi
Juhyun
J
Structural Biology Research Center, Korea Institute of Science and Technology, 39-1 Hawallkok-dong Sungbuk-ku, Seoul, South Korea.
Kim
Eunice E
EE
Park
Young In
YI
Han
Ye Sun
YS
D000818:Animals; D003001:Cloning, Molecular; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D004372:Ducks; D015684:Gene Products, pol / Q000302:isolation & purification; D005814:Genes, Viral; D005822:Genetic Vectors; D015611:Hepatitis B Virus, Duck / Q000201:enzymology*; D006515:Hepatitis B virus / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D010843:Pichia / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D013698:Templates, Genetic
D015684:Gene Products, pol; C066027:P protein, Hepatitis B virus; D011993:Recombinant Fusion Proteins; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase; D016914:Ribonuclease H