| PCR-SSCP method was used to study the OBR and PPAR genes that might be correlated with fat traits in SiChuan White Goose, XuPu Goose and Tong-Ting Grass Goose. We detected the genes' SNPs and deposited their partial sequences in GenBank. Furthermore, the frequencies of gene and genotype, genetic polymorphism and homology were analyzed in a breed and in different breeds. On the foundation of the fat traits measurement in different goose breeds, the genetic effects between SNPs genotype and fat traits of OBR and PPAR genes in goose were analyzed and the predominant genotype were screened. Consequently, the positive ESTs which significantly correlate with lipidosis trait of goose were screened by Delta Differential Display Reverse Transcription PCR (DDRT-PCR) method and the high homologue relative genes were found by cloning, sequencing and homology analysis. All the positive ESTs are submitted in GenBank database. The main results as followed.1. There were 3 genotypes in DNA fragment of OBR gene in XuPu goose and SiChuan White goose. A mutation of A→G was taken place in 56bp of this gene, which led to variation of amino acid sequence. We submitted the gene fragment to GenBank and its accession number was DQ346681. The results of BLAST showed that the gene fragment has 100% identification with turkey and chicken and 96% with human, monkey, swine, horse and dog. Moreover, the gene frequency and genotype frequency have significant differences in different breeds.2. SNPs genotypes of OBR significantly affected the abdomen fat weight (AFW), percentage of abdomen fat (AFP) and very low density lipoprotein (VLDL) concentration of blood plasma, but there were distinct differences in the two breeds. BB genotype was the predominant genotype which decreased the belly fat weight, belly fat ratio and VLDL concentration. And the contribute percentage was above 29% to XuPu goose and 20% to SiChuan White goose. However, AA genotype was the predominant genotype that increased the AFW, AFP and VLDL concentration and the contribute percentage was over 8%. In XuPu goose, the main effect index(MEI) of AFW, BFR and VLDL concentration were 44.466, 16.797 and 21.284 respectively. While in SiChuan White goose, the MEI of AFW, AFP, VLDL concentration and sebum thickness (ST) were 9.638, 8.442, 21.112 and 10.685 respectively. So the OBR gene might be the main effect gene of goose fat traits.3. There was a SNP of PPARαgene in Dong-Ting Grass goose. But it didn't modify amino acid sequence. The sequence of DNA fragment of PPARαhas 95% identification with Domestiemuscory duck, domesticated duck and mouse, and the genotypes of the SNPs affected AFW and AFP significantly. The AFW of genotype AA and BB were significantly less than that of AB and the AFP of genotype BB was significantly less than that of AB. So genotype BB was the predominant genotype that decreased the AFW and AFP. Its contribution rate was above 8%, but genotype AB brought the positive direct contribution percentage of 5.263% to AFP. The MEI of PPARαwas 12.11 in AFW and 17.31 in AFP. Thereby, it was supposed that PPARαgene was the main effect gene of goose fat traits or it linked the major gene that control the abdomen fat traits.4. Eleven differential expression positive fragments related to lipidosis in different tissue were screened by Delta DDRT-PCR method. After cloned and sequenced, the. 11 ESTs were submitted to GenBank and its accession numbers were EY199060~ EY199070. After BLAST and alignment analysis, one of the EST had 94% homology with leptin (ob) gene and another EST had 95% homology with P2RY5 gene. These results showed that the two genes had obviously association with lipidosis traits of goose. |
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