Analysis Of Sequence, Expression Patterns, And Polymorphism Of Porcine PGM1 Gene

Phosphoglucomutasel(PGM1) is up-regulated in Yorkshire muscles according to proteome analysis of porcine Longissimus Doris between Meishan and Yorkshire, Thus it is a good candidate gene affecting the pork quality. The isozymes of PGM are central to carbohydrate metabolism. PGM1 has also achieved prominence as a key marker in genetic linkage mapping and in forensic science. The aim of this work was to establish foundation for evolution research, to seek better SNPs for MAS and to investigate the potential function of 3'UTR of PGM1 on post-transcriptional level. In our research, methods of molecular biology and bioinformatics were used to study the PGM1 gene and the results are as follows:(1) Here we have isolated only one cDNA clone type corresponded to the ubiquitous transcript of PGM1 in porcine skeletal muscles, CDS is 1,689bp and encodes 562 amino acids, which is 99% similarity with the ubiquitous transcript of human PGM1. Putative amino acid sequence of Sus scrofa was closer to B.taurus PGM1 than H.sapiens on the basis of phylogenic tree among 14 selected species. These sequences spanned animal kingdom, plant and prokaryote.(2) It is implied that pig PGM1 has at least two different products since PGM1 was separated into two points in albumin gel by 2D-E during the previous studies. However, no missense mutation was detected for the diversity of swine PGM1. We suggested that the PGM1 have disparity in post-translational modification.(3) The samesense mutations A420G and C462T of which genotypes and allelic frequencies were analyzed in CDS and mutational site G1693A in 3'-UTR of PGM1 are confirmed by using PCR-RFLP and PCR-SSCP respectively. For the former two SNPs, the frequencies of the'A' and'C allele were dominant in Chinese indigenous breeds.(4) The results of correlation analyses between genotype and traits, which were performed by GLM in 284 pigs of the'Yorkshire×Meishan'F2 resoure population, showed that C462T has significant correlation with lean meat percentage(P<0.05). Chi-square test indicated that no deviation from HWE was observed in Landrace(Xc2<χ20.05(df))and Yorkshire(χc2<χ20.05(df)) population.(5) We have also studied the temporal and spatial expression profile of the PGM superfamily except PGM4 by semiquantitative RT-PCR and quantitative real-time PCR(qRT-PCR). PGM1 has higher expression in longissimus Dorsi, the lowest in Semitendinosus. We speculate that the total enzymatic activity of PGM in longissimus Dor si come from PGM1.In addition, the complete 3'-UTR, which is possibly combined with miRNAs inhibiting the translation of swine PGM1 gene, was successfully inserted into the multiple cloning site(MCS) of pEGFP-C1 vector for the further study on post-transcriptional regulation.