mRNADecayofPorcineGSTM2Gene：猪GSTM2基因的mRNA衰变.pdfVIP

ISSN 1672-9145 Acta Biochimica et Biophysica Sinica 2007, 39(8): 560–566 CN 31-1940/Q Cloning, Sequence Analysis and Identification of a Nonsense Mutation-mediated mRNA Decay of Porcine GSTM2 Gene Jingshu HUANG, Yuanzhu XIONG*, Changyan DENG, Bo ZUO, Dequan XU, Minggang LEI, and Siwen JIANG Key Laboratory of Swine Genetics and Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China Abstract The glutathione S-transferase mu 2 gene ( GSTM2) encodes a GST functioning in the elimination of electrophilic compounds and the regulation of cell growth. In this study, the sequence of porcine GSTM2 gene that contains the complete sequence encoding a protein of 218 amino acids was cloned. The deduced amino acid sequence shared 76%, 78% and 76% identity with that of human, mouse and rat, respectively. mRNA expression analysis showed that the porcine GSTM2 gene was expressed at a high level in liver and testis, at a medium level in longissimus dorsi muscle, adipose tissue, spleen and lung, at a low level in kidney, and at a very low level in heart and embryo. A nonsense mutation (CGA→TGA) resulted from C27T substitution in the fifth exon to produce a premature translation termination codon was identified, and it was discovered that nonsense-mediated mRNA decay might have an effect on the regulation of porcine GSTM2 gene expression. This polymorphism was analyzed in Large White, Landrace, Meishan and Qingping pig populations using the Taq I-polymerase chain reaction-restriction fragment length polymorphism method. The result showed that allele C had a higher frequency than allele T in each population. Keywords GSTM2; pig; NMD; gene expression; PCR-RFLP Glutathione S-transferase (GST) is biotransformation or reducing