›› 2004, Vol. 22 ›› Issue (5): 14-305.

• 实验报道 • Previous Articles     Next Articles

Molecular Cloning of a Protein Antigen Gene of Cysticercus cellulosae

LIU Dian wu,LI Wen ling,ZHANG Li mei
  

  1. Department of Epidemiology,Hebei Medical University,Shijiazhuang 050017,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2004-10-30 Published:2004-10-30

Abstract:  Objective To immunoscreen one protein antigen gene from a cDNA library of Cysticercus cellulosae . Methods A cDNA library of C. cellulosae was constructed after cDNA was synthesized, and immunoscreened using rabbit anti C. cellulosae polyclone antibody. The gene structure and its possible function were analyzed by comparing with the sequences available in the GenBank, after the insert of positive clone was subcloned to pBluescript SK plasmid and the nucleotide sequence of the cDNA was determined by dideoxynucleotide chain termination method using a Taq DyeDeoxy Terminator Cycle Sequencing Kit. The amino acid sequence was deduced from nucleotide sequence using GENETYX software. Homological search of the nucleotide sequences was done using BLAST in GenBank. Results and Conclusion A cDNA clone of 1 320bp was isolated. The clone contained one open reading frame composed of 1 260bp encoding 420 amino acids, in which two potential glycosylation sites were found. The partial nucleotide sequence of the gene fragment showed high homology with the essential spectrin gene of Caenorhabditis elegans and the erythrocyte surface antigen gene of Plasmodium falciparum , when the gene fragment was homologically analyzed in GenBank.

Key words: Cysticercus cellulosae, Molecular cloning, cDNA library, Sequencing, Homology analysis