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Nucleotide Sequence Polymorphism within Exon 1 of Aquaporin 7 Gene and Its Association with Semen Quality in Murrah Buffaloes

Ragini Kumari K. P. Ramesha Rakesh Kumar Beena Sinha P. Divya
Vol 8(2), 137-145

The present study was conducted on Murrah bulls (n= 69) with the objectives of identifying single nucleotide polymorphisms (SNPs) in targeted regions (exon 1 and its flanking intronic region) of AQP7 gene and analyzing their association with semen quality traits in Murrah bulls. Genomic DNA was isolated by using high salt method from blood of Murrah bulls. Targeted region of 459 bp of AQP7 gene was amplified using polymerase chain reaction (PCR). The PCR products were further subjected to single strand conformation polymorphism analysis (SSCP). PCR-SSCP analysis revealed three unique band patterns by silver staining of the polyacrylamide gels. BioEdit software (version 7.2) were used for detecting SNPs. Comparative sequence analysis of AQP7 gene in Murrah bulls with Bos taurus (Ensemble Ref Seq: ENSBTAG00000020105) revealed eleven nucleotide change, out of which three SNP were observed among Murrah bulls. Among these all, pattern 2 of exons 1 have significantly higher mean sperm concentration (p<0.01). Association analysis of exon 1 indicated that hypo-osmotic swelling reactivity in frozen semen was significantly influenced (P<0.05). These findings may suggest that there is genetic variation at exon 1 of AQP7 gene for better sperm quality which would serve as potential genetic marker(s) for semen quality traits in buffalo.

Keywords : AQP7 Gene Murrah Buffalo PCR-SSCP SNP

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