Cloning, Mapping, SNP Detection, And Association Analysis With Production Traits Of The Porcine FBXO32 Gene And It's Transcriptional Factor Genes

The ubiquitin-proteasome parthway (UPP) is one of the most efficient protein catabolism pathways in eukaryotic cells. It plays important roles in a number of biological processes including cell cycle, muscle atrophy, antigen triggered cell apoptosis and etc., especially in upstream and downstream regulation by dispelling activator or suppressor in different pathways in the cell. The ubiquitin protein ligase E3 mediated by F-box protein can specially recognize different substrates, and can regulate UPP.In this study, FBXO32 gene and its transcriptional factor genes (FoxO1, FoxO3a and FoxO4) related to development and growth of skeletal muscle in the UPP pathway were selected as candidate genes for investigating associations with production traits. The cloning, mapping, expression, SNP detection and association analysis of the genotypes with growth and carcass traits were conducted for these genes. The markers developed in this study could be potential markers for marker assisted selection. The results are as follows:1. The pig specific primers were designed from the pig EST contig obtained by assembling EST from GenBank according to the corresponding human cDNA. PCR products of four genes (FBXO32, FoxO1, FoxO3a and FoxO4) were successfully amplified, cloned, and sequenced. The sequences have been deposited in GenBank database, the accession numbers are: DQ288235 {FBXO32), AY995235 (FBXO32), DQ243692(FoxO1), DQ243691 (FoxO3a), DQ288236 (FoxO3a) and DQ243693 (FoxO4).2. IMpRH and SCHP were employed to determine the chromosomal locations of the four genes. The FOXO32 gene was localized to 4p15, with close linkage to SW835 (LOD=11. 49), S0301 (LOD=8. 98) and S0001 (LOD=5. 93). The FoxO1 gene was localized to 11p11-15, with close linkage to SW1632(LOD=3. 5). The FoxO3a gene was localized to 1p13, with close linkage to SW301(LOT=14. 72) and SW781 (LOD=10. 08). The FoxO4 gene was localized to xq13, with close linkage to SW1835 (LOD=5. 9).3. RT-PCR was performed to determine the expression profile of porcine FBXO32, FoxO1, FoxO3a and FoxO4 in 13 different tissues including heart, liver, spleen, lung, kidney, skeletal muscle, fat, tongue, small intestine, stomach, embryo, ovary and endometrium from the adult Landrace pigs which has been pregnant for 90 days. The results showed that these genes are expressed in most of the tissues.