T/A CLONING AND SEQUENCE ANALYSIS OF PHOSPHOGLYCERATE KINASE GENE OF SCHISTOSOMA JAPONICUM

Objective To clone and sequence the partial gene of Schistosoma japonicum phosphoglycerate kinase( SjPGK ), and to provide a new candidated antigen for preventing schistosomiasis japonica. Methods A pair of primers were designed and synthesized according to the cDNA sequence of S. mansoni phosphoglycerate kinase (SmPGK ) gene. The gene fragment of SjPGK was amplified and isolated from the total RNA of S. japonicum by reversetranscription polymerase chain reaction(RTPCR). The gene fragment was cloned into the T/A cloning vector of pMD18T, the positive clones were acquired and identified with restrictive enzymes and PCR amplification. After being sequenced with DNA autosequence analysis instrument, the cDNA sequence of SjPGK was searched for homology identity with NCBI Blast program. Results The gene encoding SjPGK was obtained and isolated by RTPCR. The fragment of SjPGK was about 830 bp. The cDNA sequence of the phosphoglycerate kinase was highly homology between S. mansoni and S. japonicum. The identity of nucleotide sequence was 85% and score 672; The identity of amino sequence was 94% and score 473. Conclusion The partial gene of encoding SjPGK is successfully cloned into the cloning vector of pMD18T, which provided the basis for further study.