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Expression of the X Gene of Hepatitis B Virus
Author(s) -
Pugh John C.,
Weber Christian,
Houston Heather,
Murray Kenneth
Publication year - 1986
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890200305
Subject(s) - virology , gene , biology , hepatitis b virus , microbiology and biotechnology , gene product , plasmid , open reading frame , herpes simplex virus , genome , escherichia coli , capsid , dna , virus , gene expression , peptide sequence , genetics
The hepatitis B virus (HBV) genome carries an open reading frame of 462 bases, the X region, but the corresponding protein has yet to be identified as a natural product. In rodent cells cotransformed with the thymidine kinase gene of herpes simplex virus and HBV DNA, however, Gough [1983] identified a mRNA that hybridises uniquely with the X region of the HBV genome. A large fragment of the X region was inserted into plasmid pCL19LDY‐T in order to produce, in Escherichia coli , the X gene product, HBxAg, as a polypeptide fused to the N‐terminal part of the phage þcro gene product. Antisera raised against this fused polypeptide gave positive immunofluorescence reactions with the transformed rodent cells. This provides direct evidence for the expression of the HBxAg gene in eukaryotic cells transformed with HBV DNA. The approach used here should be generally applicable.