Subcellular Location of ARRDC5 Gene in Banna Mini-pig Inbred Line

Purpose The function of the arrestin domain-containing protein 5 (ARRDC5) was unclear up to now. Our previous research indicated that ARRDC5 might be associated with the fecundity of boars. The aim of this study was to clone the porcine ARRDC5 gene and analyze its subcellular location.

Methods The ARRDC5 sequences of allied species of pig were obtained from NCBI and compared by online BLAST to obtain the expressed sequence tag (EST). Subsequently we used silicon cloning strategy to gain the coding sequence and a few flanking sequences of ARRDC5. Then, the eukaryotic expression vector pEGFP-C1-ARRDC5 was constructed by Digestion-Ligation and transfected into the swine testicle (ST) cells with liposome, detection of protein expression by inverted fluorescence microscopy.

Result The cDNA length of Banna mini-pig inbred line (BMI) ARRDC5 was 1 045 bp that included an open reading frame of 1 014 bp coding a protein of 337 amino acids. The BMI ARRDC5 protein sequence shared high homology with that of cattle and human. We successfully constructed the expression vector pEGFP-C1-ARRDC5, and the green fluorescent was mostly limited in the cytoplasm of ST cells, which showed that the gene products of ARRDC5 in eukaryocytes were mainly located in the cytoplasm. The cell nuclei are also expressed in some cells.

Conclusion These results will provide a basis for the study of ARRDC5 function in swine.