Cloning Of 5' Flanking Region Of Goose PRL Gene,Association Of SNPs With Egg Production

PRL which secreted from anterior pituitary gland has extremely broad spectrum of biological role.In avian,PRL is especially crucial for reproduction traits such as broodiness and thus for egg production.Stubborn broodiness of local avian species mainly attributes to their low egg production.The level of PRL in avian blood plasma was regarded as one of the factor in intriguing broodiness.In this study,the WanXi geese(n=200),a Chinese local breed,and Rhine geese (n=80)were used to detect SNPs in the PRL subunit gene.PRL gene is as to candidate gene,by genome comparison,one pair of primers was designed using Primer Premier5.0 and DnaStar programme in cloning of 5' flanldng region of goose prolactin gene.Considering the association of PRL with goose reproductivity,the PRL gene was selected as a candidate to screen the nucleotide variation by means of PCR-SSCP protocol and analyze the association of polymorphisms with early egg reproduction.The partial of 5' flanking region of goose prolactin gene was cloned(GenBank accession number:EF190486)by the method of Genomic comparison.Homologic analysis suggested that the homology of 5' flanking region of goose prolactin with chicken PRL gene was 73%,and 78%,69%with Ceylon jungle fowl and Japan quail PRL gene respectively.There are three SNPs in the 5' flanldng region of goose prolactin,A-401G,G-268A and T-266A.The putative binding transcriptional proteins of 5'-flank region were predicted using TFSEARCH ver1.3 on line.The result revealed that G-268A and T-266A lead to-changes of transcription factor.They caused to lose two binding sites for GATA-1 and GATA-2.Genetic analysis suggested that,among Wanxi-White goose population,two allele frequencies detected by P2 primer were 0.7275 and 0.2725 respectively,allele frequencies detected by E2 were 0.6688and 0.3312 respectively.In Rhine goose population,allele frequencies detected by P2 and E2 were 0.665,0.0875 and 0.6125, 0.3875 respectively.?~2 fitness test suggest that the population were in the state of Hardy-Weinberg disequilibrium at the two site.Association analysis of early egg production traits and PRL gene polymorphisms in geese showed that SNPs in 5'flanking region influenced egg production traits of Wanxi-White geese.The egg number of genotype AA were lower(P＜0.05)than that of AB and BB.In exon 2 site,the weight of first egg of genotype CC were lower (P＜0.05)than that of CD and DD in Wanxi-White geese.The weight of first egg of genotype EE were lower(P＜0.05)than that of EFand FF in Rhine geese...