[Expression of DNA fragment encoding for immunodominant epitopes of 150kd phosphoprotein of human cytomegalovirus in Escherichia coli].
HVYP fragment, which contains the DNA encoding region for immunodominant epitopes of 150 kd phosphoprotein (pp150) of human cytomegalovirus, was isolated from recombinant plasmid pHCY1 and the inserted into the PstI site of E. coli expression vector pUC8. The resulted recombinant plasmid pUHVYP1 with correct orientation and in-frame was transformed into E. coli JM107. Induced by isopropyl-beta-D-thiogalactoside (IPTG), the transformants produced HVYP fusion protein with molecular weight of about 44-45 kd, as determined by 12% SDS-PAGE. These results from Western blot and ELISA showed that the expressed protein could be recognized by the specific anti-HCMV IgG and IgM antibodies of patients' serum. MAC-ELISA further proved that the fusion protein was a potential antigen for diagnostic reagents.
Wang
Z
Z
Department of Biotechnology, Sichuan University.
Tang
Z
Z
Zhang
Y
Y
D003001:Cloning, Molecular; D003062:Codon; D003587:Cytomegalovirus / Q000235:genetics*; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014760:Viral Fusion Proteins / Q000276:immunology
D003062:Codon; D000939:Epitopes; D010750:Phosphoproteins; D014760:Viral Fusion Proteins