| DNA molecular marker technology , such as PCR-RFLP, PCR-SSCP, RMAPD (Random-microsatellite amplified polymorphic DNA), DNA sequence analysis, microsatellite marker, were applied to evaluate the effects of genotypes of six candidates genes (CSN1S2 gene, CSN3 gene, GH gene, IGFBP3 gene, POU1F1 gene, FSHR gene) in Xinong Saanen Dairy goat; And PCR-RFLP and PCR-SSCP was used to detect genetic diversity of five goat breeds (including Xinong Saanen dairy goat, Guanzhong dairy goat, Shaanan White goat, Angora goat and Boer goat) . The main goal were to construct the foundation of making good use of economic traits (milk performance, litter sizes and body sizes) in Xinong Saanen dairy goat and protecting breed resources, so that molecular theory about breeding in Xinong Saanen dairy goat and constructing useful database in five goat breeds were provided primary. The results were as follows:1. Research on PCR-RFLP in Xinong Saanen dairy goatCSN1S2 gene: (1) In 69 Xinong Saanen dairy goats population which was at Hardy-weinberg equilibrium, the frequencies of allele F/N for CSN1S2 gene loci were 0.087/0.913; (2) Allele F was always bound with lower milk yield, such as genotype FF individuals showed lower milk yield than that of genotype NN in average milk yield and 2nd milk lactation (P<0.05, P<0.01). (3) CSN1S2 gene locked with FecB gene affected litter sizes because different genotypes for CSN1S2 gene loci had significant difference in 1st litter sizes and 4th litter sizes (P<0.05,P<0.01). Besides, there was also significant difference among genotypes in body sizes (P<0.05). CSN3 gene: No polymorphisms for CSN3-HaeIII gene loci was detected; The locus of CSN3-HindIII and CSN3-TaqI had no significant effect on milk performance; It was verified firstly in this trial that CSN3, CSN1S2, CSN1S1 and CSN2 gene locks with FecB gene because litter sizes of CSN3-TaqI gene loci had significant difference among different genotypes in 1st litter sizes and 2nd litter sizes(P<0.05, P<0.01). Hence CSN3, CSN1S2, CSN1S1 and CSN2 genes can be regarded as candidate genes for effective molecular marker about litter sizes.By the way, the polymorphic loci also affect some body sizes (P<0.05).IGFBP3 gene: The results about restriction nucleauase HaeIII,Alw26 and TaqI digesting IGFBP3 gene demonstrated singular genotype. With DNA sequence analysis for DNA sequence FSHRexon10 of from bovine heterozygote and Xinong Saanen goat , no changes in HaeIII restiction cleavage site (form 299th to 302nd ) resulted in no polymorphism of IGFBP3 gene. While obvious polymorphisms of IGFBP3 gene digested with HaeIII appeared among goat, sheep, buffalo and bovine.POU1F1 gene: In Sixty-nine individuals of Xinong Saanen dairy goat and sixty-two individuals of Guanzhong dairy goat populations, there were no polymorphism at HinfI cleavage site in POU1F1 gene. 2. The polymorphisms of SNP loci detected by PCR-SSCP in Xinong Saanen dairy goat and its relationship with economic traits No polymorphism was discovered in FSHRexon10 gene loci; while GHexonII gene showed polymorphism in Xinong Saanen dairy goat population which was at Hardy-weinberg didn't associated with milk yield (P<0.05), and GHexonV gene concluded genotype AB,AC, BB, BC which showed significantly difference in milk performance in 3rd milk yield , 4th milk yield , 5th milk yield , 6th milk yield and 7th milk yield and average milk yield . Moreover, the different genotypes in GHexonII and GHexonV gene loci affected significantly litter sizes in 3rd litters sizes, 5th litter sizes and 7th litter sizes. To analyze SNPs of GHexonV gene by DNA sequence analysis in this population, there was transition or transversion in 11 places. As far as difference between goat and bovine in FSHRexon10 was considered, the result of DNA sequence analysis showed amino acid change for Gln→His appears in 54th loci for goat.3. Microsatellite MarkerIn this trial, two anonymous microsatellite primer from fish were amplified in Xinong Saanen goat genomic DNA and no polymorphism was detected.4. RMAPD... |
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