Synthesis of a recombinant DNA-derived HBV e antigen and its application in diagnosis.
The gene coding for HBV e antigen (HBeAg) and core antigen (HBeAg) are located in the C region of the HBV genome. In this paper 1.8 kb BamHI-EcoRI DNA fragment carrying HBc gene from adw2 HBV DNA was digested with Bal31 and HapII, a series of fragment with different length were produced and inserted into pUR222 and pUC18 vectors to construct various recombinant plasmids. Two new recombinants able to direct high level synthesis of HBeAg in E. coli were obtained. Recombinant HBeAg was then purified by DEAE chromatography and affinity chromatography. The preparation obtained were identified by electrophoresis and immunochemical analysis. Molecular weight of the recombinant HBeAg determined by SDS-PAGE was about 38 kd. It is considered to be a dimer of two fused antigen molecules in size of 19 kd. Bacterial extracts prepared from cells harboring one of the constructed plasmids have been used successfully as a diagnostic reagent for the detection of HBeAg and anti-HBe in human sera by ELISA.
Wu
G H
GH
Institute of Hepatology, Beijing Army General Hospital.
Huang
Y X
YX
Ni
L P
LP
Duan
L X
LX
Jia
K M
KM
D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D004926:Escherichia coli; D005069:Evaluation Studies as Topic; D006509:Hepatitis B / Q000175:diagnosis*; D006513:Hepatitis B e Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D010957:Plasmids; D015183:Restriction Mapping; D012680:Sensitivity and Specificity
D004274:DNA, Recombinant; D004279:DNA, Viral; D000939:Epitopes; D006513:Hepatitis B e Antigens