Cloning And Expression Of CC1 Antigen Of Cysticercus Cellulosae For The Serodiagnosis Of Cysticercosis

Objective: cC1 gene was amplified from Cysticercus cellulosae。The recombinantprotein was used to generate a new detection for cysticercosis. Methods: Specificoligonucleotide primers were designed and synthesized. Total RNA was isolated fromCysticercus cellulosae using Trizol reagent. Coding region gene of cC1 wasamplified by RT-PCR technique. The PCR product was cloned into pGEM-T vector.Then the cC1 was subcloned into pET28a expression vector and the recombinantplasmid was transformed into E. coli BL21. The fusion expression of cC1 was inducedby IPTG. The expression was subjected to SDS-PAGE followed by identification withWestern blotting. We used this recombinant protein as antigen to detect antibodiesagainst Cysticercus cellulosae for diagnosis of cysticercosis by means of ELISA.Results: The coding gene of cC1 was specifically amplified by RT-PCR. The size ofthe cC1 DNA fragment in pGEM-T-cC1 and pET28a-cC1 digested with BamHⅠandXhoⅠwas identical to the length of the PCR generated products. DNA sequencingconfirmed that the cC1 gene had an open reading frame of 1044bp with an approximatemolecular weight of 38kDa and was homologous to cC1 gene in GenBank (AF147955).The recombinant plasmid was highly expressed in E.coli BL21. SDS-PAGEelectrophoresis showed a band with molecular weight of 40kDa as expected. The fusioncC1 expression was verified by Western blotting using the sera of patients withcysticercosis. The primary detection of antibodies to cC1 in the patients withcysticercosis showed a promising sensitivity and specificity with ELISA. Conclusion:The cC1 gene of Cysticercus cellulosae was successfully cloned and expressed and theresults demonstrated that recombinant cC1 antigen had a high sensitivity and specificitywhen used for the immunodiagnosis of cysticercosis. It provides the basis for furtherstudy on serodiagnosis and, potentialy, on the vaccine development for cysticercosis.