Cloning, Mapping, Snp Detection And Association Analysis With Production Traits Of The Porcine Gfat1 Gene And Six Genes Of Slc2a Gene Family

The objective of this thesis was to isolate GFAT1 genes and six genes of SLC2Agene family on pig glouse metabolism passway using information derived fromcomparative mapping and pig EST database. The cloning, mapping, expression, SNPdetection and association analysis of the genotypes with growth and carcass traits wereconducted for these genes. The markers developed in this study could be potentialmarkers for marker assisted selection. The results are as follows:1. The pig specific primers were designed from the pig EST contig obtained byassembling EST from GenBank according to the corresponding human eDNA. PCRproducts of seven genes (GFAT1,SLC2A2,SLC2A4,SLC2A5,SLC2A8 and SLC2A12)were successfully amplified, cloned, and sequenced. The sequences have been depositedin GenBank database, the accession numbers are: EF011104 (SLC2A2), EF012367(SLC2A3), EF012368 (SLC2A5), EF012369 (SLC2A8), EF012370 (SLC2A12).2. SCHP and IMpRH were employed to determine the chromosomal locations of thesix genes. The GFAT1 gene was localized to 3q21-q27, with close linkage to SW828(LOD=12.39). The SLC2A2 gene was localized to 13q23-(1/2 q41), with close linkage toS0084 (LOD=4.29). The SLC2A3 gene was localized to 5q25, with close linkage toSW963 (LOD=14.46). The SLC2A5 gene was localized to 6q22-q23, with close linkage toSW1355 (LOD=6.65). The SLC2A8 gene was localized to 1q28-q213, with close linkageto SW705 (LOD=4.32). The SLC2A12 gene was localized to 1p24-p25, with close linkageto SW1851 (LOD=6.22).3. RT-PCR was performed to determine the expression profile of porcine GFAT1 in 9different tissues including heart, liver, spleen, lung, kidney, skeletal muscle, fat, smallintestine and stomach from the adult Landrace which has been pregnant for 90 days andadult Tongcheng pigs, and in 6 different tissues including skeletal muscle, heart, liver,spleen, lung, kidney from small LandrancexLarge White which has been born for 4days.The results showed that GFAT1 gene is expressed in most of the tissues, but it isselectively expressed in skeletal muscle and heart. RT-PCR was performed to determinethe expression profile of porcine SLC2A4 in 9 different tissues including heart, liver,spleen, lung, kidney, skeletal muscle, fat, small intestine and stomach from adultTongcheng pigs. The results showed that SLC2A4 gene is expressed in most of the tissues,in the tissue of skeletal muscle and heart, it is expressed relatively higher.4. One SNP was detected in the intron 8 of GFAT1 and the exon 4 of SLC2A4respectively. The mutation in exon 4 of SLC2A4 did not cause amino acid change. For the two SNPs, PCR-RFLP was performed to detect the genotypes in different porcinebreeds.The results showed that the GFAT1 SNP does not segregate in domestic breeds(Tongcheng), it showed polymorphism in YLT. SLC2A4 SNP is polymorphic in mostbreeds.5. The intron 8 MvaI—RFLP polymorphism of GFAT1 and the exon 4 HinlI—RFLPpolymorphism of SLC2A4 were detected and analyzed in five pig populations includingYongcheng, Landrace, Large White, Landrace×(Large White×Yongcheng) and LargeWhite×(Landrace×Tongcheng). The results showed that different genotyps of GFAT1SNP showed significant differecnes in carcass straight length and Backfat at the rump (depth)(P＜0.05). The different genotypes of SLC2A4 SNP had significant differences in Muscleshearing force (P＜0.05).