巴马小型猪OCA2基因序列、组织表达分析及载体构建研究

doi:10.11843/j.issn.0366-6964.2016.03.004

Abstract

Abstract:

In order to establish scientific basis for farm animal models associated with coat color mechanism and human diseases in OCA2 gene，we carried out this experiment.The exons 20-24 of porcine OCA2 gene(NC_010457.4) in absence in GenBank were cloned and mapped.The mapping result was confirmed at the genomic and cDNA levels using PCR amplification and Sanger sequencing simultaneously.Bioinformatics analysis was performed for OCA2 gene.Polymorphic loci of all exons for OCA2 gene were screened in Guangxi Bama Miniature pigs.Meanwhile，the mRNA expression in different tissues was detected by real-time fluorescence quantitative PCR(Q-PCR).A full-length skin cDNA was used as a template to construct recombinant expression plasmid，and then XhoⅠand SacⅡrestriction enzyme double digestion was carried out.We mapped and cloned the porcine OCA2 gene in GenBank successfully.A total of 10 SNP loci were found in all exons，including 7 synonymous mutations and 3 missense mutations（R124H，G509D，R573H）.Porcine OCA2 gene was widely expressed in various tissues.The expression in lung，brain and cerebellum was at relatively higher level compared to that in spleen，stomach，caecum and skin，which exhibited moderate expression level，and the expression level in heart and liver was lower.pEGFP-N1-OCA2 recombinant plasmid was constructed successfully after confirmation of restriction enzyme double digestion and sequencing.This study explored porcine OCA2 gene sequence information and mRNA expression preliminarily，and recombinant expression plasmid of OCA2 gene was constructed，which provide the foundation for the study of disease model and gene biological function in future.

CLC Number:

S828
S813.3

HONG Qian-long，ZHANG Ying，CAO Chun-wei，WANG Xian-long，ZHAO Jian-guo. Analysis of Sequence，Tissue Expression and Vector Construction of OCA2 Gene in Bama Miniature Pigs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 439-448.

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Analysis of Sequence，Tissue Expression and Vector Construction of OCA2 Gene in Bama Miniature Pigs

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