Cloning, sequence characterization, and tissue expression profile analysis of three novel porcine genes—RHOB, RHOG, and PRAF1

Abstract

The complete coding sequences of three porcine genes—Rho-related GTP-binding proteins RHOB and RHOG and Prenylated Rab acceptor protein 1 (PRAF1) were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) based on the sequence information of the mouse or other mammals and referenced highly homologous pig ESTs. The nucleotide sequences of these three genes revealed that porcine RHOB gene encodes a protein of 196 amino acids that contains the conserved putative RhoA-like domain and has high homology with the RHOB precursor of three species—human, rat, and mouse (100%).The porcine RHOG gene encodes a protein of 191 amino acids that contains the conserved putative RhoG domain and has high homology with the RhoG precursor (RHOG) of three species—human, mouse, and Cricetus cricetus (98%). The porcine PRAF1 gene encodes a protein of 185 amino acids that contains the conserved putative PRA1 domain and has high homology with the PRAF1 of five species—dog (97%), cattle (97%), human (96%), rat (95%), and mouse (95%). The tissue expression analysis indicated swine RHOB gene was moderately expressed in lung; weakly in fat, spleen, and kidney; and almost not expressed in small intestine, large intestine, liver, and muscle. The swine RHOG gene was overexpressed in small intestine, large intestine, liver, and muscle; moderately expressed in kidney; weakly in spleen; and almost not expressed in fat and lung. The swine PRAF1 gene was overexpressed in fat and spleen, moderately in lung and kidney, weakly in small intestine and large intestine, and almost not expressed in liver and muscle. Our experiment established the primary foundation for further research on these three swine genes.