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SNV and haplotype analysis reveals new CSRP1 variants associated with growth and carcass traits
Authors:Hua He  Xiao-lin Liu  Hui-lin ZhangJing Yang  Fu-biao NiuZhi-xiong Li  Yu LiuLing Chen
Affiliation:College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China
Abstract:The cysteine and glycine-rich protein 1 and 2 genes (CSRP1 and CSRP2) are an effective growth factor in promoting skeletal muscle growth in vitro and vivo. However, in cattle, the information on the CSRP1 and CSRP2 genes is very limited. The aim of this study was to examine the association of the CSRP1 and CSRP2 variants with growth and carcass traits in cattle breeds. Three single nucleotide variants (SNVs) were identified within the bovine CSRP1 gene, whereas CSRP2 gene has not detected any SNVs, using DNA pooled sequencing, PCR-RFLP, and forced PCR-RFLP methods. These SNVs include g. 801T>C (Intron 2), g. 46T>C (Exon 3) and g. 99C>G (Intron 3). Besides, we also investigated haplotype frequencies and linkage disequilibrium (LD) coefficients for three SNVs in all study populations. LD and haplotype structure of CSRP1 were different between breeds. The result of haplotype analysis demonstrated eight haplotype present in QC (Qinchuan) and one haplotype in CH (Chinese Holstein). Only haplotype 1 (TTC), shared by all two populations, comprised 10.74% and 100.00%, of all haplotypes observed in QC and CH, respectively. Haplotype 5 (CTC) had the highest haplotype frequencies in QC (30.98%) and haplotype 1 had the highest haplotype frequencies in CH (100.00%). The statistical analyses indicated that one single SNV and 19 combined haplotypes were significantly or highly significantly associated with growth and carcass traits in the QC cattle population (P < 0.05 or P < 0.01). Quantitative real-time PCR (qRT-PCR) analyses showed that the bovine CSRP1 and CSRP2 genes were widely expressed in many tissues. The results of this study suggest that the CSRP1 gene possibly is a strong candidate gene that affects growth and carcass traits in the Chinese beef cattle breeding.
Keywords:SNV, single nucleotide variants   CSRP1, cysteine and glycine-rich protein 1 gene   CSRP2, cysteine and glycine-rich protein 2 gene   PCR-RFLP, polymerase chain reaction-restriction fragment length polymorphism   forced PCR-RFLP, forced polymerase chain reaction-restriction fragment length polymorphism   LD, linkage disequilibrium   QC, Qinchuan   CH, Chinese Holstein   qRT-PCR, quantitative real-time PCR   CRPs, cysteine-rich proteins   SMCs, smooth muscle cells   Ho, gene homozygosity   He, gene heterozygosity   Ne, effective allele numbers   PIC, polymorphism information content   LSD, least significant difference   GLM, general linear model   Hap, haplotype   Fat, abdominal fat
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