Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (18): 3639-3648.doi: 10.3864/j.issn.0578-1752.2016.18.018

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Sequence Cloning, Tissue Expression Profile and Polymorphism of VRTN Gene in Suhuai Pig

WU Yi-chen1, Du Xing1, LI Ping-hua1,2, WU Yan3, WANG Jun-shun3, LIU Hong-lin1, Li Qi-fa1   

  1. 1 College of Animal Science and Technology, Nanjing Agriculture University, Nanjing 210095
    2 Huaian Academy of Nanjing Agricultural University, Huaian 223005, Jiangsu
    3Huaiyin Pig Breeding Farm, Huaian 223322, Jiangsu
  • Received:2015-12-26 Online:2016-09-16 Published:2016-09-16

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Abstract: 【Objective】The aim of this study is to obtain the coding region sequence of Suhuai VRTN gene, identify its characteristics and expression patterns, analyze the polymorphism of VRTN ins291 in Suhuai pig population. 【Method】The coding sequence of Suhuai pig VRTN gene was isolated by cloning and sequencing. The BioEdit 7.0 software was used to analyze the molecular characteristics and physicochemical properties of Suhuai pig VRTN. Nucleotide and amino acid sequences alignment was performed using Clustal W. Genomic organization and chromosomal locations were investigated by comparing the cDNA and corresponding genomic sequence (UCSC server and NCBI database). Motif analysis was performed using the online programs SMART, and protein tertiary structure was predicted by CPHmodels server. Tissue samples of heart, liver, spleen, lung, kidney, hypothalamus, ovary and skeleton muscle were obtained from adult female Suhuai pigs (n=3). Total RNA was extracted using a Trizol kit, and the expression patterns of VRTN gene of Suhuai pig were analyzed by RT-PCR. The ear samples were obtained from adult female Suhuai pigs (n=106), and genomic DNA was extracted by a conventional phenol-chloroform extraction method. PCR-Gel electrophoresis was performed to analyze the polymorphism of VRTN ins291. 【Result】The full length of the coding sequences of Suhuai pig VRTN gene is 2097 bp, which shared similarities of 84.98%, 74.53%, 85.84% and 86.45% to human, mouse, cattle and sheep, respectively, but only 54.42% to chicken. Genomic structure analysis showed that pig VRTN gene includes 2 exons and 1 intron, and is located on chromosome 7. The Suhuai pig VRTN gene encodes 698 amino acid residues, which shared similarities of 85.55%, 70.07%, 86.20% and 86.06% to human, mouse, cattle and sheep, respectively, but only 51.17% to chicken, indicated that VRTN is conserved in the evolution process of mammalian. Protein structure analysis found that VRTN protein of Suhuai pig contains several typical domains including HTH domain, and comprised 6 α-helices, 5 β-sheets and many random coils. Real-time PCR analysis revealed that the VRTN gene is expressed in heart, liver, spleen, lung, kidney, hypothalamus, ovary and skeleton muscle of Suhuai pig, suggesting that VRTN is widely expressed in various tissues of pig. Genotyping ins291 of VRTN gene in 106 Suhuai pigswas done, and three genotypes were detected. In Suhuai pigs, the frequencies of wt/wt, wt/Q and Q/Q genotypes were 0.717, 0.198 and 0.085. The frequencies of high vertebrate number of Q allele was 0.184. The polymorphic information content of VRTN ins291 in Suhuai pigs was 0.331. The heterozygosity of VRTN ins291 in Suhuai pigs was 0.40. 【Conclusion】 The coding region of VRTN gene was isolated, and its expression showed no tissue-specificity. The presence of high number of vertebrate Q allele in Suhuai pig population was confirmed.

Key words: Suhuai pig, VRTN gene, cloning, tissue expression profile, polymorphism

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