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 aa(104-124), 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 alpha helix-beta sheet, with the key sites such as substrate binding 
region, active sites, Mg(2+)-binding sites closely located at the center. The 
protein contains a potential nuclear localization sequence aa(190-199) and 
several linear B cell epitopes and CTL T cell epitopes, of which the outside 
epitope aa(49-57) and inside epitope aa(228-236) are facultative T cell and B 
cell epitope, and the linear B cell epitope aa(206-213) contains the active 
center site Glu(210), 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.