Molecular cloning and expression of a novel proteasome β1 gene from Clonorchis sinensis

  Objective   To explore the proteasome B1 gene(CsPSMB1)from Clonor chis sinensis.To provide the functional study foundation of the CsPSMB1.

  Methods   From a cDNA library of Clonorchis sinensis adult worm, the cDNA inserts in pBluescriptⅡSK(+) were identified by large-scale sequencing and a novel CsPSMB1 gene were identified.A pair specific oligonucleotide primers were designed and syntheses according to the coding sequence of CsPSMB1.The coding region of CsPSMB1 was amplified by PCR and then cloned into the vector of pGEX-4T-1.The recombinant plamid was identified by PCR and endonuclease digestion then transformed into BL21(DE3).The fusion protein was expression after IPTG induction for 3 hours and identified by SDS-PAGE.

  Results   A novel CsPSMB1 gene was identified from Clonorchis sinensis adult worm cDNA library.The 749 bp cDNA encoded a putative protein of 217 amino acids with a predicted molecular mass of 24kD.The putative protein contained a single proteasome domain and founded to be 50% identical to the PSMB1 of homo sapiens.A recombinant plamid pGEX-4T-1-CsPSMB1 was constructed and expressed the GST fusion protein by IPTG induce.

  Conclusion   A novel CsPSMB1 gene was identified from Clonorchis sinensis adult worm cDNA library.A recombinant plamid pGEX-4T-1-CsPSMB1 was constructed and expressed the GST fusion protein by IPTG induce.