›› 2003, Vol. 21 ›› Issue (3): 5-146.

• 论著 • Previous Articles     Next Articles

Cloning and Identification of the Gene Fragments of Paragonimus westermani

LING Jia-jian,HOU Min,LIU Jian-nan,ZHANG Zi-hao,ZHANG Yao-juan
  

  1. Department of Pathogenic Biology and Immunology,Nanjing Medical University,Nanjing 210029
  • Received:1900-01-01 Revised:1900-01-01 Online:2003-06-30 Published:2003-06-30

Abstract:  Objective To screen and identify the recombinants from the cDNA library of the adult Paragonimus west-ermani (PwA) for immunodiagnosis and immunoprophylaxis. Methods PwA cDNA library was screened with the PwA antigen immunized rabbit sera(IRS) pre-absorbed by the extract of E. coli XLl-Blue. The recombinants from positive clones were amplified by PCR, sequenced and cut off by KpnI/BarnHI and, then sub-cloned into pRESETB vector. The fusion protein was expressed,analysed by SDS-PAGE and identified by Western blotting with immune rabbit serum against worm antigen of Paragonimus westerrnani. Results The inserted cDNA fragment from the positive clone Pw-2 was about 800 bp, which contained an open reading frame(ORF) encoding Pw pre-procathepsin L belonging to cysteinase family. Expression product of Pw-2 was a fusion protein of 32 kDa, which can be recognized by immune rabbit serum against worm antigen of Paragonimus 晈esterrnani. Conclusion A recombinant plasmid Pw-2 encodes Pw pre-procathepsin L is constructed.

Key words: Paragonimus westermani, cDNA library, recombinant protein