Expression of multivalent pre-S1 antigen of hepatitis B virus in Escherichia coli (synthetic oligodeoxynucleotides, surface antigens, recombinant DNA, fusion protein, beta-galactosidase).
The nucleotide sequence encoding 30 amino acids (aa) of the pre-S1 envelope region of the human hepatitis B virus has been constructed from twenty chemically synthesized oligodeoxynucleotides by simultaneous ligation. The DNA fragment containing four repeated sequences encoding the pre-S1 region (aa 20-49) has been inserted into the lacZ gene of the plasmid pWR450.1, yielding the recombinant pWX4 plasmid. The Escherichia coli DH5 strain transformed with pWX4 produces a beta-galactosidase-[-pre-S1(20-49) x 4] fusion protein. The hybrid protein containing 127 aa of repeated pre-S1 region has been isolated from Escherichia coli as inclusion bodies and purified by anion exchange chromatography. The antigenic properties of this fusion protein were confirmed by immunoblotting with pre-S1-specific monoclonal antibodies.
Sidorkiewicz
M
M
Department of Biochemistry I, Medical School, Warsaw, Poland.
Jaworski
J
J
Płucienniczak
G
G
Płucienniczak
A
A
Stpień
P
P
Kalinowska
B
B
Karwowska
S
S
Nowosławski
A
A
Uznański
B
B
Wilk
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D004274:DNA, Recombinant; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D009838:Oligodeoxyribonucleotides; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen; D001616:beta-Galactosidase