High-level production of hepatitis B viral X protein in Escherichia coli using gene II promoter of bacteriophage M13.
Region X is one of the four open reading frames (ORFs) of hepatitis B virus (HBV) and encodes a polypeptide of 154 amino acids (aa). A 584-bp BamHI-BglII fragment of the HBV DNA containing the major part of ORF X which encodes 145 aa was inserted into the BglII site within the gene II of bacteriophage M13. The insertion resulted in an in-phase gene II-X fused protein of 174 aa under the control of the gene II promoter. Cells harboring plasmids (pML alpha X.59 and pMLX.12d) derived from the above construct overproduced the 19-kDa fused protein in Escherichia coli at a level of 10%-20% of total cellular protein. The fused protein was recognized by the anti-X antibodies. This is the first demonstration of using gene II promoter of M13 to express a foreign gene efficiently.
Chen
M L
ML
Graduate Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.
Lee
Y H
YH
Lo
S J
SJ
D010582:Bacteriophage lambda / Q000235:genetics; D003090:Coliphages / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D006510:Hepatitis B Antibodies / Q000276:immunology; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
D006510:Hepatitis B Antibodies; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins