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Single nucleotide polymorphism scanning and expression of the FRZB gene in pig populations
Authors:Zhixiu Wang  Qinggang Li  Bo Zhang  Yunfeng Lu  Yuzeng Yang  Dongmei Ban  Hao Zhang
Affiliation:1. National Engineering Laboratory for Animal Breeding, China Agricultural University, No. 2 Yuanmingyuan West Rd., Beijing 100193, People''s Republic of China;2. Institute of Animal Sciences and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230031, Anhui Province, People''s Republic of China;3. School of Life Science & Technology, Nanyang Normal University, Nanyang 473061, Henan Province, People''s Republic of China
Abstract:Secreted frizzled-related protein 3 (sFRP3), encoded by the gene FRZB, is a member of the sFRP family with important roles in inhibition of the Wnt signalling pathway through competitive binding of the Wnt receptor. Here, we investigated pig FRZB as a candidate gene for growth traits and identified three polymorphic sites, an insertion (A-532B) and two SNPs (G636A and C650T) in its 5′-UTR. The genotype distributions of G636A and C650T were significantly different among mini-type indigenous (Diannan Small-ear and Tibetan), normal indigenous (Laiwu and Huai), and introduced (Large Yorkshire and Landrace) breeds. In semi-quantitative PCR expression analysis, expression of FRZB mRNA was abundant in tissues of hypophysis, longissimus dorsi muscle, and adipose tissues, and low in the heart, hypothalamus, and brain. Quantitative determination of mRNA level and protein expression analysis were corresponding. The results demonstrated that FRZB gene expression in longissimus dorsi muscle and liver tissue was significantly higher in Diannan Small-ear and Tibetan pigs than in the Large Yorkshire breed (P < 0.05); however, in back fat tissue, the expression was significantly higher in Diannan Small-ear pig than in Tibetan or Large Yorkshire breeds (P < 0.05). Given the known growth and fat characteristics of the breeds, these results indicate that FRZB expression has a negative association with muscle growth and a positive association with fat deposition. In conclusion, FRZB may be a major candidate gene for growth traits in pigs.
Keywords:SNP, single nucleotide polymorphism scanning   sFRP3, secreted frizzled-related protein 3   FRZB, frizzled motif associated with bone development   UTR, untranslated region   CRD, extracellular cysteine-rich domain   DSP, Diannan Small-ear pig   TP, Tibetan pig   LW, Laiwu pig   HP, Huai pig   LY, Large Yorkshire   LL, Landrace   LD, longissimus dorsi muscle   DNA, deoxyribonucleic acid   RNA, ribonucleic acid   PCR, Polymerase Chain Reaction   SDS-PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis   PBST, Phosphate Buffered Saline with Tween-20   HPRT, hypoxanthine phospho-ribosyl transferase   mRNA, messenger RNA   HWE, Hardy&ndash  Weinberg Equilibrium
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