[Comparison of the Chinese LJ strain structural gene with HM175, MBB, LA strains and the expression of hepatitis A virus antigen by LJ/HM175 recombinant vaccinia virus].
The nucleotide and amino acid sequences of a Chinese hepatitis A virus Long-Jia (LJ) strain were compared with that of HM175, MBB and LA strains in structural genes (nt 630-3049). The most extensive nucleotide homology was identified between LJ and MBB strains. The identity rates of nucleotide were 95.4%, 96.7% and 91.4%, respectively. Variation rates of amino acid were 0.91%, 0.91% and 2.98%, respectively. A total of 23 amino acid differences located in whole capsid region between LJ and LA strain, especially in VP1. Only 7 amino acid differences located in VP2 and VP3 between LJ and HM175/MBB strain. Restriction enzyme sites increased 10, 13 and 30 sites in 56 restriction enzymes tested, and decreases 15, 1 and 27 sites, respectively. BstE II (nt 2810) and Pvu I (nt 2013) were the peculiar sites of LJ strain. Hind III, Pst I and Sac I sites were identical among the four strains. After structural gene (nt 745-2993) of HM175 strain was replaced by LJ strain, the complete hepatitis A virus cDNA open reading frame was inserted into pJSA1175 (vaccinia virus expression vector) downstream of promoter 7.5 k. Hepatitis A virus antigen expressed was 1:16 in titer by sandwich ELISA. Band-patterns of anti-VP0, anti-VP1 and anti-VP1, 2, 3, were as same as that of HM175 strain and natural hepatitis A virus antigen by Western blot analysis.
Xia
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Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
Yi
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Y
Guo
K
K
Tian
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H
D035921:Hepatitis A Antigens / Q000235:genetics* / Q000276:immunology; D030041:Hepatitis A virus / Q000235:genetics* / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014616:Vaccinia virus / Q000235:genetics* / Q000276:immunology; D015678:Viral Structural Proteins / Q000235:genetics* / Q000276:immunology
D035921:Hepatitis A Antigens; D015678:Viral Structural Proteins