Coding Sequence Cloning, Expression and Bioinformatics Analysis of Swine Male Enhancer 1 Gene

Purpose The aim of the present study is to obtain the CDS sequence of MEA1 gene, and to get the expression profile of the MEA1 and to know the basic function of MEA1 swine protein.

Methods The specific primers were designed based on GenBank MEA1 mRNA sequences of pig and related species, and the CDS and flanking sequence of MEA1 gene were amplified from the testis tissue of Banna mini-pig inbred line (BMI). The amino acid sequence of MEA1 protein was used to carry out functional bioinformatics analysis and construct the phylogenetic tree. Real-time quantitative PCR was applied to analyse the MEA1 gene expression profiles of 15 important tissues.

Results We obtained a 525 bp CDS of BMI MEA1 gene, which encodes a peptide of 174 amino acids. Bioinformatics analysis indicated that the MEA1 protein contained a complete MEA1 domain without transmembrane region and signal peptide sequences. Its secondary structure is predominantly composed of random coil and its N-terminal is hydrophobic and C-terminal is hydrophilic. The MEA1 protein has three different modification sites. BMI MEA1 had the closest relationship with that of human and gibbon by phylogenetic analysis. This demonstrated that protein sequences are coincident with classical taxonomy. Real-time quantitative PCR in multi-tissues indicated that MEA1 gene was expressed highly in the testis tissue, and in other 14 tissues were weak.

Conclusion Our study will lay a foundation for further study of the MEA1 gene functions in pig testicular development and spermatogenesis.