猪CBR1基因不同拷贝形式克隆及其多态性分析

doi:10.3864/j.issn.0578-1752.2018.08.018

Abstract

Abstract: 【Objective】 In order to obtain and analysis the different copy forms of CBR1 gene coding region, and analysis the correlation between polymorphism and reproductive performance of different copy forms of CBR1 gene in pigs.【Method】 Based on the cDNA of the Wuzhishan pig’s testis , the different copy sequences of CBR1 gene were obtained by cloning sequencing technology. The amino acid sequences encoded by different copy forms of CBR1 gene were analyzed by DNASTAR software. Homology between different copy forms of porcine CBR1 gene, and that compared with other mammals were analyzed using DNAMAN software. The physical and chemical properties of different proteins of porcine CBR1 gene were analyzed by Protparam online software on ExPaSy. 136 Yorkshire and 47 Landrace pig ears samples were selected for DNA extraction and PCR-sequencing to explore the relationship between polymorphism of CBR1 gene and porcine reproductive performance. 【Result】 All the three different copies of CBR1 gene encoded 3 exons, and the sequence length of the coding region were 870bp, 870bp and 846bp respectively. Their 12-18th amino acid residues were GlyXXXGlyXGly, which were in accordance with the CBRs family structural characteristics. The results of homology comparison showed that the CDS sequence homology among different copies of CBR1 gene was more than 87%, and more than 82% among different mammals, indicated that the CBR1 gene of mammal was conserved in evolution process. The physical and chemical properties of 3 different copies of CBR1 were similar, all of them were unstable and belong to hydrophilic proteins, which indicated that they may play a similar role in vivo. Polymorphism analysis showed that 5 SNPs were found in copy V1, 4 SNPs were found in copy V2, and 2 SNPs were found in copy V3. In the Yorkshire pigs, the A/T mutation at the promoter upstream region 153 bp and the C/T mutation at exon3 379 bp in V1; AC insertion at the promoter upstream region 10 bp, C/T mutation at exon 1 63 bp and G/T mutation at intron1 76 bp in V2, were significantly correlated with the multiparous alive litter size, while the SNPs in Landrace pigs were not associated with reproductive performance. 【Conclusion】 Coding region sequence of 3 different copies of CBR1 gene were obtained; 5 SNPs were found significantly associated with multiparous alive litter size , suggested that CBR1 gene may be used as a valuable candidate gene for improving breeding performance of Yorkshire pigs.

Key words: swine, CBR1 gene, re-sequencing, de-novo assembly

QI ChuanXiang, XU KUI, MU YuLian, YANG ShuLin, LI Kui, WU TianWen. The Cloning of Porcine CBR1 Gene’s Different Copy Forms and Analysis of Its Polymorphism[J].Scientia Agricultura Sinica, 2018, 51(8): 1607-1616.

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The Cloning of Porcine CBR1 Gene’s Different Copy Forms and Analysis of Its Polymorphism

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