Cloning and Expression Analysis of CIDEA in Adipose Tissue and Adipocytes of Jinchuan Yak

doi:10.7668/hbnxb.20192192

Abstract

Abstract: The aim of this study was to clone the gene sequence of cell death-inducing DNA fragmentation factor 45-like effector A (CIDEA) in yak, analyze its biological characteristics, and detect the expression pattern of CIDEA in yak different tissues and during the differentiation of adipocytes. The CDS sequence of CIDEA was cloned by PCR, using Jinchuan yak subcutaneous adipose tissue cDNA as a template, and the biological characteristic was analyzed;specific primers were designed to detect the expression of CIDEA in various Jinchuan yak tissues, including lung, heart, kidney, fat and other tissue of Jinchuan yak. And primary adipocytes were isolated to analyze the expression trend of CIDEA during adipocyte differentiation. The results showed that the length of Jinchuan yak CIDEA was 696 bp, of which CDS was 684 bp, encoding 227 amino acids. CIDEA of Jinchuan yak had the highest homology to that of Bos taurus, representing that the nucleotide identity was 94.14% and the amino acid homology was 99.54%, and the lowest homology to that of Gallus gallus, representing that the nucleotide identity only was 63.38% and the amino acid homology only was 59.05%. Also, phylogenetic tree constructed by MEGA 5.0 software showed that similarity was highest between Jinchuan yak and cattle, and similarity was lowest between Jinchuan yak and chicken. Moreover, the physical and chemical properties of protein were predicted that CIDEA protein was an unstable basic hydrophilic protein without transmembrane structure and signal peptide. The secondary structures of CIDEA protein mainly consisted of 33.92% α-helix and 51.98% random coils. Real-time quantitative PCR results showed that the expression quantity of CIDEA was highest in Jinchuan yak adipose tissue and was lowest in lung. And the differentiation of adipocytes in Jinchuan yak showed an increasing trend. It is concluded that the expression of CIDEA might involve in adipose differentiation and adipose deposition in yaks.

Key words: Jinchuan yak, CIDEA, Gene cloning, Temporal-expression, Adipose tissue, Adipocyte

CLC Number:

Q78
S823

HUO Wentao, ZHOU Ting, YU Yue, YANG Qinglan, CHANG Xindan, LI Jian, XIONG Xianrong, XIONG Yan. Cloning and Expression Analysis of CIDEA in Adipose Tissue and Adipocytes of Jinchuan Yak[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 224-231. doi: 10.7668/hbnxb.20192192.

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