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Published on: December 21, 2011
[Molecular cloning and characterization of a RRas homologue gene from Trichomonas vaginalis]
Ming-yan Xu1, Yu-cai Fu, Ju-li Liu
1Department of Parasitology, Shantou University Medical College, Shantou 515041, China.
Objective:
To clone and characterize a RRas-like gene from Trichomonas vaginalis for studying cellular signal transduction pathways in the organism.
Methods:
A cDNA clone, which showed homology with RRas proteins of human being, was isolated and sequenced from a cDNA expression library of T. vaginalis. The genomic DNA corresponding to the cDNA sequence was amplified using PCR technique and sequenced. Sequence analysis was performed using BLASTP, RPS-BLAST and Clustal W programs. Phylogenetic tree was constructed and bootstrapped with 1050 replicates using the software MEGA3.
Results:
The cDNA sequence showed a length of 705 bp with an open reading frame of 615 bp. The deduced amino acid sequence from the open reading frame possesses 205 residuals. Sequencing of the PCR product of genomic DNA revealed that the genomic DNA sequence encompassing the putative 5'-ATG and 3'-stop codons was identical to the cDNA sequence. Sequence analysis demonstrated that this gene was most homologous to the RRas members of Homo sapiens and Mus musculus (both having 51% identity and 70% similarity), and the amino acid sequence contains highly conserved GTP-binding domains and a fully conserved effector domain of human RRas member. Phylogenetic analysis showed that TvRRas clustered with RAS oncoprotein branch and RRAS branch of human.
Conclusion:
The encoding protein probably belongs to a RRas family of T. vaginalis.
