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- Title
Reverse vaccinology approach identify an Echinococcus granulosus tegumental membrane protein enolase as vaccine candidate.
- Authors
Wenjia Gan; Guoxiong Zhao; Hongxu Xu; Weihua Wu; Wuying Du; Jiang Huang; Xinbing Yu; Xuchu Hu
- Abstract
Applying reverse vaccinology strategy, we employed a sequence encoding an enolase from Taenia asiatica to search its homolog in the expression sequence tag (EST) database of Echinococcus granulosus and found two EST sequences (Access number: CN653186 and CN649593) of a clone Eg_PSGRS_13B09 from E. granulosus protoscolex full-length cDNA library, which are responding for the 5' and 3' partial cds of E. granulosus enolase, respectively. Primers are designed according to the 5' end and 3'end of the putative encoding sequence to amplify the genomic DNA of E. granulosus strain isolated from sheep in Qinghai province of China by polymerase chain reaction (PCR). A sole product of 1,449 bp in length was obtained, which contains two little introns of 78 bp and 69 bp, respectively. The introns were excised by unsymmetrical PCR with combined flank sequences of introns as primers. The structural, functional, and immunological characteristics of putative amino acid sequence were predicted by bioinformatics analysis. The complete coding sequence was predicted to encode 433 residues and contain a transmembrane region aa104-124, with the N terminus outside and C terminus inside. The inside part is quite the functional domain. SWISS-MODEL modulated its 3D structure as a barrel which constitutes of alternatively arranged α helix-β sheet, with the key sites such as substrate binding region, active sites, Mg2+-binding sites closely located at the center. The protein contains a potential nuclear localization sequence aa190-199 and several linear B cell epitopes and CTL T cell epitopes, of which the outside epitope aa49-57 and inside epitope aa228-236 are facultative T cell and B cell epitope, and the linear B cell epitope aa206-213 contains the active center site Glu210, suggesting the putative protein is a potential membrane with strong immunogenicity. The complete cds was expressed in Escherichia coli, and the recombinant protein can be recognized by the serum from patient infected with E. granulosus. Reverse vaccinology process identified E. granulosus tegumental membrane protein enolase as vaccine candidate.
- Subjects
ECHINOCOCCUS granulosus; ECHINOCOCCUS; ENOLASE; TAENIA; MEMBRANE proteins
- Publication
Parasitology Research, 2010, Vol 106, Issue 4, p873
- ISSN
0932-0113
- Publication type
Article
- DOI
10.1007/s00436-010-1729-x