title: Detection of functional polymorphisms influencing the promoter activity of the SAA2 gene and their association with milk production traits in Chinese Holstein cows.
abstract: Our previous RNA sequencing experiment showed that the serum amyloid A2 (SAA2) gene was one of the most promising candidates for milk protein and fat traits in dairy cattle. The SAA2 gene encodes an apolipoprotein related to high-density lipoproteins. To further validate its genetic effects, genotype-phenotype associations were performed in this study. Through resequencing of the entire coding region and the 5'-regulatory region of the SAA2 gene using pooled DNA of 12 unrelated sires, one novel 3-bp insertion-deletion and five previously reported SNPs were detected. These identified SNPs were genotyped and tested for association with five milk production-related traits in 717 Chinese Holstein cows. After Bonferroni correction for multiple t-tests, five of them were found to be statistically significant for milk yield, fat yield and protein yield (P < 0.0001~0.0053). Haplotype-based association analysis revealed a similar effect on fat yield and protein yield (P = 0.0005, P = 0.0032 respectively). Then, using luciferase report assay, the regulatory effect of the three SNPs located in the promoter region (c.-22G>A; c.17G>C; c.114G>A) was evaluated on transcriptional activity. In HEK-293 cell lines, we found that constructs GCG and AGG showed higher luciferase activity compared with GCA (P < 0.01, P < 0.01 respectively). Meanwhile, the prediction of the putative differential transcription factor binding site revealed that c.17G>C and c.114G>A caused the alteration in the transcription factor. Overall, the findings presented here provide the first evidence for associations of the SAA2 gene with milk fat and protein traits, which appears to be a key candidate for milk production traits in dairy cattle.
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: S
authors.initials: S
authors.lastname: Yang
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: C
authors.initials: C
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Xie
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: X
authors.initials: X
authors.lastname: Cui
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: J
authors.initials: J
authors.lastname: Wei
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Yu
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture, National Engineering Laboratory of Animal Breeding, China Agricultural University, Beijing, 100193, China.
authors.forename: D
authors.initials: D
authors.lastname: Sun
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D005260:Female; D056726:Genetic Association Studies; D057809:HEK293 Cells; D006239:Haplotypes; D006801:Humans; D054643:INDEL Mutation; D015810:Linkage Disequilibrium; D008892:Milk; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D000685:Serum Amyloid A Protein / Q000235:genetics*
chemical_list: D000685:Serum Amyloid A Protein
keywords: SNP; dairy cattle; genetic effect; luciferase; transcriptional activity