Study Of PRLR Gene Single Nucleotide Polymorphism And Copy Number Variation Of Duck

In this research, Brown Tsaiya Ducks were used as experimental materials, PCR-SSCP was applied to detect and analyze the association of polymorphism of PRLR gene exon 10 and their combined genotypes with production traits.The recombined plasmids were constructed successfully, containing of target gene and reference gene, respectively. The amplification efficiency of target gene and reference gene showed no difference. Then used 2-△△Ct method to estimate the copy number of PRLR gene and analyze the association of copy number variation of PRLR gene and their combined genotypes with production traits.The results were as follows:(1) The sequencing results showed that there were 1 mutation in PRLR gene enxon 10 and two genotypes were identified. The frequency of allele B and b were 0.787 2 and 0.212 8, the heterozygosity was 0.335 0, polymorphism information content was 0.278 9, which belongs to an intermediate polymorphism. χ2 test suggested that the population was out the state of Hardy-Weinberg equilibrium at the two sites. Association analysis of early production traits showed that the individuals of genotype Bb were significant higher than that of genotype BB in birth weight and egg weight at 300 days (P<0.05). The individuals of genotype Bb were extremely significant higher than that of genotype BB in shell weight.(2) Primers of PRLR and Ldh-B were designed and recombinant plasmids were cloned as standard templates. Standard curves for primers were established using qPCR with 10-fold dilution templates. The standard curve of PRLR and Ldh-B gained from qPCR showed that the correlation coefficient were 0.991 and 0.990, amplification efficiency were 111.68% and 108.429%, the slope of standard curve were -3.070 and -3.135, respectively.The amplification efficiency of primers of target gene and reference gene were approximately the same,2-△△Ct method would be used for quantitative analysis of PRLR gene.(3) In the 94 Brown Tsaiya Ducks, five kinds of copy number variation were identified. Association analysis of early production traits showed that the individuals with 2 copy number were significant higher than that of 1 copy number and 5 copy number for shell thickness (P<0.05), the individuals with 3 copy number were significant lower than that of 1 copy number,2 copy number and 5 copy number for egg shape index (p<0.05).