PurposeThe aim of this study was to obtain the function of STRA8 gene in Banna mini-pig inbred line (BMI) and lay a foundation for further study of the gene in spermatogenesis role.
MethodReferring to STRA8 JQ965783 and EST sequences of pig downloaded from GenBank, we designed specific primers and amplified Banna mini-pig inbred line (BMI) STRA8 gene. The physicochemical properties and functional bioinformatics of STRA8 protein were analysed including the secondary structure, protein domain, hydrophobicity, transmembrane structure, signal peptide, subcellular localization, phosphorylation sites and protein function. The STRA8 amino acid sequences of other species were searched and downloaded for constructing phylogenetic trees. The mRNA transcription expression levels in BMI 15 tissues was detected by real-time quantitative PCR.
ResultThe result obtained the BMI STRA8 full length sequence 1 316 bp, including coding sequence 1 101 bp, with a protein of 366 amino acids, which has been submitted to GenBank with mRNA accession number KU70562. The molecular weight (Mw) and the isoelectric point (pI) of this protein were 41.06 ku and 4.52, respectively. The most of secondary structure of STRA8 protein was alpha helix, which accounted for 52.19%. The BMI STRA8 protein contained a protein functional domain, an HLH structure domain and a curly spiral region in the amino terminal, two low-complexity regions in the carboxyl terminal. Hydrophobic analysis showed that the both N-terminus and C-terminus are hydrophilic without transmembrane structure and signal peptide sequence. The probability of the protein located in the nucleus is 56.5%. The STRA8 has 34 phosphorylation sites. Phylogenetic result demonstrated that BMI has the highest similarity with sheep and water buffalo. The mRNAs of STRA8 were differential expression in the BMI 15 tissues and there was the highest expression in the testes.
ConclusionThis study will provide a reference for studying STRA8 gene function in pig spermatogenesis.
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