Study On The Polymorphism Of FSHβ, FSHR And The Association With Reproduction Traits In Wusuli Raccoon Dog

Wusuli Raccoon Dog is a valuable animal whose skin is covered with fur. It became famous for tenacity of fur and unique guard hair. In recent years, it could provide good opportunities of development for fur-bearing animals with the market of fur recoveried. Therefore, the economic benefits of wusuli raccoon dog could be effectively improved by increasing the litter size and account of livestock on hand. The differences of genetic phenotypes between wusuli raccoon dogs were brought by traditional breeding techniques, and the differences of reproductive traits of populations were the largest, which brought great economic losses to feed economic animal. Therefore, it's important to improve litter size with the modern molecular biology techniques for the wusuli raccoon dog breeding.Follicle-stimulating hormone beta subunit (FSHβ)and its receptor (FSHR) gene were viewed as the candidate gene. In the studies in livestock, FSHβand its receptors have been identified as major genes of reproductive traits.In our study, the experimental groups were the wusuli raccoon dogs feeding in the farms of the wusuli raccoon. dogs in Jiangbei of Harbin. The single nucleotide polymorphisms (SNPs) of gene FSHβand FSHR gene were detected with the techniques of single strand conformation polymorphism (PCR-SSCP) and DNA sequencing methods. The correlation between gene polymorphism and reproductive traits analysis was analyzed. The main results are as follows: 1. FSHβgene was cloned and sequenced in the Wusuli raccoon dog. It's full-length was 3246bp. This sequences included the first 1 to 3 exons, intron 1 and 2, and the partial 5'upstream regulatory sequence of FSHβ. It has been released to the Genbank, and it's accession number is HQ 385979.2. Two polymorphic sites of FSHβgene were detected in the Wusuli raccoon dog with PCR-SSCP technique. They are respectively located in G192A of 5'regulatory region and A2393G in exon 3. The corresponding frequencies of genotype and allele of the genetic polymorphisms were calculated. The correlative analysis of SNPs and the reproductive traits showed that the CC,CD and DD genotypes resulting from the A2393G mutation has some influence on litter size of the Wusuli raccoon dog (P<0.2). The DD genotype has a greater effect on the litter size than CC genotype in the Wusuli raccoon dog. There were not significant differences among litter size of the AA, BB and AB genotype caused by the G192A mutation.3. Exon 10 and the 5'sequence of FSHR genes were cloned and sequenced in the Wusuli raccoon dog. The length of exon 10 sequence was 1398bp and it has been released to the Genbank. It's accession number was HQ 385979. The length of 5'sequence of FSHR was 757bp which included regulatory sequences and exon 1 sequence. It's Genbank accession number was HQ385977.4. The comparison of 5'sequence and exon 10 of FSHR gene between wild and domesticated Wusuli raccoon dog showed that there were multiple mutations in the FSHR gene in the wild Wusuli raccoon dog. These mutations included 46 to 49 TTAG insertion sites,452～453AC insert,448～451 missing ACAC, and 9 SNPs in the sequence sites, while only one SNPs in the domesticated raccoon dogs. Exon 10 was highly conserved, only 3 polymorphisms was found in exon10 of FSHR in wild Wusuli raccoon dog.5. Two polymorphic sites of FSHR gene were detected in the Wusuli raccoon with PCR-SSCP technique. They are located in the T133A of 5'regulatory region the and T623C of exon 1.The corresponding frequencies of genotype and allele of these two polymorphisms were calculated. The correlation analysis showed that there were not significant differences between the litter size of these two SNPs in the Wusuli raccoon dog.