Cloning, expression and display of the PreS domain of hepatitis B virus on filamentous bacteriophage M13.
The PreS domain of hepatitis B virus (HBV) is believed to be involved in virion assembly and attachment to a hepatocyte receptor during infection. In order to study the functions of this region, we fused it to the g3p protein of bacteriophage M13 that allows the fusion protein to be displayed at the tip of the filament. The fusion protein was detected by the anti-E tag antibody on a Western blot. The polypeptide in a soluble form was produced by transfecting a non-suppressor E. coli host cell with the recombinant phagemid. The soluble protein was detected in cytoplasm, in the periplasmic space and also in the medium. The functional display of the PreS domain would provide an alternative means to study its interactions with the nuleocapsid and hepatocytes.
Department of Biochemistry and Microbiology, Faculty of Science and Environmental Studies, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia.
W L
WL
Kok
K
K
Yusoff
S
S
Nathan
W S
WS
Tan
D017104:Bacteriophage M13 / Q000235:genetics*; D015153:Blotting, Western; D003001:Cloning, Molecular; D003470:Culture Media; D003593:Cytoplasm / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006514:Hepatitis B Surface Antigens / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014169:Transformation, Bacterial
D003470:Culture Media; D006514:Hepatitis B Surface Antigens; D011993:Recombinant Fusion Proteins