Cloning and sequencing of the gene encoding variant-specific surface antigen from Giardia lamblia.
OBJECTIVE
To clone and sequence variant-specific surface antigen gene from Giardia lamblia isolate SUCH/89/BTMR/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 published 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 nucleotide and the amino acid levels.


CONCLUSIONS
The full length variant-specific surface antigen gene from the isolate of G. lamblia has the common characteristics with other previously identified VSPs.
Li
Ya-Jie
YJ
Department of Pathogenic Biology, Medical College of Dalian University, Dalian 116622, China. Liyj@dlu.edu.cn
Teng
Mei-Jun
MJ
Lun
Yong-Zhi
YZ
Li
Da
D
Zhang
Yong-Qing
YQ
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D016829:Giardia lamblia / Q000235:genetics*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D016054:DNA, Protozoan; D015800:Protozoan Proteins