[Construction of plasmids expressing hepatitis E virus antigen fragments and their combinations].
OBJECTIVE
Several different regions of the HEV antigen were expressed in Escherichia coli and their immunological characteristics were simply evaluated.


METHODS
Three coding sequences of immunodominant antigenic regions in structural protein of the hepatitis E virus (ORF2.1: 6287-6403nt, ORF2.2: 6743-7126nt, ORF3: full length of ORF3) were amplified by RT-PCR from the fecal supernatants of macaques experimentally infected with HEV. These three fragments were inserted into the pThioHisC expression vector separately or fused in line by (Gly)n short arms. After identified by DNA sequence analysis, these six recombinant plasmids were transformed into Escherichia coli, and immunology characteristics of expressed antigen fragments were evaluated by Western blotting.


RESULTS
All six genes were successfully expressed in Escherichia coli. Western blotting assay showed that the recombinant proteins specifically reacted with the serum antibody from hepatitis E patient in various degrees.


CONCLUSIONS
Artificial antigen based on strung epitopes provides a promising strategy for detection and prevention of HEV infection.
Tang
H
H
Department of Molecular Biology, Institute of Medical Biology, CAMS and PUMC, Kunming 650118, China.
Ma
Y
Y
Li
C
C
Du
R
R
Le
G
G
Zhuang
J
J
Liu
Y
Y
Sun
M
M
Dai
C
C
D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D016752:Hepatitis E virus / Q000235:genetics / Q000276:immunology*; D010957:Plasmids / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics*; D011995:Recombination, Genetic
D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins