Bioinformatic analysis,cloning and expression of Schistosoma japonicum SCP/TAPS gene family

The Vespula vulgaris Wasp Venom Allergen 5(Vv Vesv5,GenBank accession No.AAA30333) protein sequence was used in a tBLASTn search of S.japonicum ESTs.Detail bioinformatic analysis were conducted and an interesting gene were cloned and expressed to investigate the role in the life cycle of S.japonicum.After PCR confirmation of the sequences from parasite-derived cDNA,AAW24579 was selected for further study and named S.japonicum Venom allergen-like transcript(SjVAL2).The coding region of SjVAL2 was amplified by RT-PCR using specific primers and cloned into prokaryotic expression vector pET-28a(+),then expressed in E.coli BL21(DE3).The purified recombinant protein was detected by Western Blotting.There were at least 17 different genes among S.japonicum SCP/TAPS gene family.Multiple sequence alignment indicated that they could be divided into three groups.The coding region of SjVAL2 was amplified by RT-PCR using S.japonicum adults cDNA as template.PCR,double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid was successfully constructed.The recombinant protein could be highly expressed in E.coli BL21(DE3).Western blotting indicated that rSjVAL2 could be recognized by serum from S.japonicum naturally infected mouse.SCP/TAPS gene family might play an important role in the life cycle of S.japonicum and its detail biological function need further studies.