›› 2008, Vol. 26 ›› Issue (3): 9-202.

• 实验研究 • Previous Articles     Next Articles

Cloning and Sequencing of the Gene Encoding Variant-Specific Surface Antigen from Giardia lamblia

LI Ya-jie*, TENG Mei-jun, LUN Yong-zhi, LI Da, ZHANG Yong-qing
  

  1. Department of Pathogenic Biology,Medical College of Dalian University,Dalian 116622,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-06-30 Published:2008-06-30

Abstract: Objective To clone and sequence variant-specific surface antigen gene from Giardia lamblia isolate SUCH/89/BTMRI/2(C2) derived from human in China. Methods Total genomic DNA of G. lamblia was extracted and a full-length variant-specific surface antigen gene fragment was amplified by polymerase chain reaction (PCR). The PCR product was cloned into pMD19-T simple-vector, transformed into an Escherichia coli JM109 strain and then sequenced.The sequence analysis for cloned fragment was finished by Vector NTI 9.0 software for the homology of Giardia variant-specific surface antigen gene to that of sequences publishend in GenBank. Results The full-length variant-specific surface antigen gene fragment from G. lamblia was found to be 2 142 bp, encoding a 713 amino acid polypeptide and contained a single open reading frame (ORF). The deduced polypeptide sequence was rich in cysteine (11.8 mol%), most of which occurred with in 29 copies of the 4-amino acid CXXC motif, one GGCY-tetrapeptide motifs and three NXS consensus N-linked glycosylation sites. This polypeptide was also rich in threonine (10.2 mol%), glycine (12.1 mol%) and alanine (10.1 mol%). Like other previously identified VSPs, it contained a highly conserved hydrophobic C-terminal region. The homology of G. lamblia SUCH/89/BTMRI/2(C2) variant-specific surface antigen gene to that of sequence(TSA417) published in GenBank was 99% both at the nueleotide and the amino acid levels. Conclusion The full length variant-specific surface antigen gene from the isolate of G. lamblia has the common characteristics with other previously identified VSPs.

Key words: Giardia lamblia, Variant-specific surface antigen, Sequence, Homology, Clone