VASA is a member of the DEAD-box protein family that plays an indispensable role 
in mammalian spermatogenesis, particularly during meiosis. In the present study, 
we isolated, sequenced, and characterized VASA gene in buffalo testis. Here, we 
demonstrated that VASA mRNA is expressed as multiple isoforms and uses four 
alternative transcriptional start sites (TSSs) and four different 
polyadenylation sites. The TSSs identified by 5'-RNA ligase-mediated rapid 
amplification of cDNA ends (RLM-5'-RACE) were positioned at 48, 53, 85, and 88 
nucleotides upstream relative to the translation initiation codon. 3'-RACE 
experiment revealed the presence of tandem polyadenylation signals, which lead 
to the expression of at least four different 3'-untranslated regions (209, 233, 
239 and 605 nucleotides). The full-length coding region of VASA was 2190 bp, 
which encodes a 729 amino acid (aa) protein containing nine consensus regions of 
the DEAD box protein family. VASA variants are highly expressed in testis of 
adult buffalo. We found five variants, one full length VASA (729 aa) and four 
splice variants VASA 2, 4, 5, 6 (683, 685, 679, 703 aa). The expression level of 
VASA 1 was significantly higher than rest of all (P < 0.05) except VASA 6. The 
relative ratio for VASA 1:2:4:5:6 was 100:1.0:1.6:0.9:48.