Identification and comparative analysis of full-length of porcine TRAF4 mRNA sequence

Abstract

Tumor necrosis factor receptor associated factor 4 (TRAF4) is a member of the TNF receptor associated factor family, which involved in cancer metastasis, production of reactive oxygen species, and cell polarity. During mouse embryonic development, TRAF4 was expressed from 8.5 to 13.5 days of post-coitum. Hence TRAF4 at tight junction of cell membrane is related with epithelial to mesenchymal transition, it may be essential in early embryogenesis to start of cell differenciation. However, little is known about characteristics and functions of TRAF4 in pig, specifically preimplantation embryonic development in porcine. This study was performed to investigate an isolation and identification of TRAF4 mRNA sequences in pig as the first step to understand relevant characteristics and functions. To analyze the full-length of TRAF4 mRNA, it was carried out rapid amplification of cDNA ends with total RNA from pK15 cells. Using 5’ and 3’ TRAF4 specific primers, three PCR products were obtained and applied for sequencing analyses. Consequently, 2030 bp nucleotide sequences were analyzed, seeming to putative full-length mRNA of TRAF4. The coding region of porcine TRAF4 was shown to 93 and 90 % homologies of human and mouse, respectively. The sequences were encoded to 470 amino acids of which the sequences were different from 12 and 7 amino acids of human and mouse, respectively. It was found that porcine TRAF4 contained GTWRGS domain at the C-terminal region, known to be in only TRAF4 among other TRAF family members in human. Based on these sequencing results, it can be postulated that the porcine TRAF4 mRNA sequences may provide an important information for an initial study to understand characterizations and functions of TRAF4 during porcine preimplantation embryonic development.