Molecular characterization and expression analysis of the NLR family CARD containing five transcripts in the pig.

Abstract

The NOD-like receptor (NLR) family caspase recruitment domain-containing 5 (NLRC5) is one of the newly discovered and largest NLR family members. The NLRC5 has recently received extensive attention because of its important role in regulating innate and adaptive immune responses. The NLRC5 in many vertebrates, such as humans, mice, cattle, and horses, has already been proven and studied. However, the NLRC5 gene characteristics of pigs remain unclear. Thus, we completely cloned the NLRC5 cDNA sequence of the pig using the rapid amplification of cDNA ends(RACE) technology. A characteristic and tissue expression analysis was also conducted on the pig sequence. The sequence analysis showed that the complete cDNA sequence of the NLRC5 of the pig is 6638 bp, and the open reading frame is 5538 bp which encoded 1846 amino acids. The protein prediction analysis indicates that the overall performance of the NLRC5 protein of the pig is hydrophilic and possesses a typical nucleotide binding and oligomerization domain(NBD) and 20 leucine-rich repeats(LRRs). The homology analysis result indicates that the NLRC5 transcript in pigs is highly homologous to cattle, sheep, macaques, and humans, and accounts for around 80%. The genetic evolutionary tree analysis shows that the NLRC5 transcript in pigs has the closest evolutionary relationship with cattle and sheep. Further tissue expression analysis shows that immune organ systems (e.g., lymph node and spleen) and mucosa organs (e.g., intestinal lymph node, stomach, and lungs) possess high expressions with NLRC5 mRNA. The result of this study indicates that the NLRC5 transcript in pigs is relatively conservative among mammals and may play a vital role in immune reaction, which provides a basis for further studies on the NLRC5 function in the pig immune system and the role in comparative immunity.