Cloning, Identification and Tissue mRNA Relative Expression Level Analysis of Porcine Selenoprotein P Gene

TANG Jiayong, ZHAO Hua, HE Aihua, ZHOU Jichang, JIA Gang, LIU Guangmang, CHEN Xiaoling, CAI Jingyi

doi:10.3969/j.issn.1006-267x.2016.03.027

Chinese Journal of Animal Nutrition >

2016 , Vol. 28 >Issue 3: 858 - 863

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.03.027

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Cloning, Identification and Tissue mRNA Relative Expression Level Analysis of Porcine Selenoprotein P Gene

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1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China;
2. Molecular Biology Lab, Shenzhen Center for chronic Disease Control, Shenzhen 518020, China

Received date: 2015-09-20

Online published: 2016-03-14

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Abstract

The objective of this experiment was to clone and identify porcine selenoprotein P gene (Sepp1), and investigate its mRNA relative expression level in porcine tissues for further study of its roles using pig models. Total RNA was extracted from pig liver for 3'-RACE with primer designed according to a Sepp1-like EST sequence, and assembled with the EST sequence. We further investigated the mRNA relative expression level in 9 tissues of pigs by the qPCR technology. The results showed as follows: 1) 1 707 bp cDNA fragment of the Sepp1 containing the open reading frame (ORF) till to its poly (A) tail was isolated, and a 2 109 bp full length cDNA was acquired. The sequence of porcine Sepp1 was submitted to NCBI GenBank with accession number of EF113596.2. The 1 170 bp ORF share an 83.72% identity to that of human, while their amino acid sequences had 75.64% identity. The ORF of porcine Sepp1 encode 390 amino acids, which has 14 selenocysteine (Sec) encoded by TGA codon, the Sec residues located at positions of 59, 267, 286, 309, 311, 327, 339, 352, 354, 361, 376, 378, 385 and 387 from N-to C-terminal of the mature SelP. 2) The Sepp1 mRNA was wildly expressed in various tissues and exhibited the highest mRNA distribution in liver, followed by thyroid, kidney, testis, hypothalamus, spleen, pituitary, heart and muscle. In conclusion, the porcine Sepp1 is cloned and identified; also its mRNA distribution in porcine tissues is investigated in present study, which offers an effective alternative to further study its biology roles using pig models.

Key words： pig; Sepp1; clone; RT-qPCR

Cite this article

TANG Jiayong, ZHAO Hua, HE Aihua, ZHOU Jichang, JIA Gang, LIU Guangmang, CHEN Xiaoling, CAI Jingyi . Cloning, Identification and Tissue mRNA Relative Expression Level Analysis of Porcine Selenoprotein P Gene[J]. Chinese Journal of Animal Nutrition, 2016 , 28(3) : 858 -863 . DOI: 10.3969/j.issn.1006-267x.2016.03.027

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