藏猪CDKN1B基因克隆、生物信息学及表达分析

doi:10.16431/j.cnki.1671-7236.2023.06.003

Abstract

Abstract: 【Objective】 The purpose of this study was to explore the role of cyclin-dependent kinase inhibitor 1B (CDKN1B) gene in hypoxia adaptation.【Method】 The CDS region of CDKN1B gene of Tibetan pigs was cloned and sequenced, and its sequence was compared and phylogenetic tree was constructed.The bioinformatics analysis of CDKN1B protein was carried out using online software.The relative expression of CDKN1B gene mRNA and protein in heart, kidney and lung tissues of Tibetan pigs and Yorkshire pigs were detected by Real-time quantitative PCR and Western blotting.【Result】 The CDS region of CDKN1B gene in Tibetan pigs was 597 bp, and was 100% similar to Sus scrofa.It was the closest to Sus scrofa and the farthest to Barchydanioreriovar.CDKN1B protein was composed of 198 amino acids and had 30 phosphorylation sites, including 17 serine sites, 9 threonine sites and 4 tyrosine sites, and the amino acid instability index (Ⅱ) was 66.53.The protein was hydrophilic with a maximum hydrophobicity of 1.089 and a minimum of -2.867. The secondary structure of CDKN1B protein in Tibetan pigs was composed of random coil (73.23%), alpha helix (15.66%), extended chain (10.10%) and beta turn (1.01%).The prediction of the tertiary structure was consistent with that of the secondary structure.The protein was a non-transmembrane protein and non-secretory protein, which was mainly located in the nucleus, accounting for about 82.6%, 8.7% in the cytoplasm, and 4.3% in the mitochondria and cytoskeleton, respectively.The correlation between CDKN1B protein and CDK2, CCND1 and CDK4 was as high as 99.9%.The expression of CDKN1B mRNA in Tibetan pigs and Yorkshire pigs was the same as that of protein, which was the highest in heart, followed by kidney and lung.The mRNA expression of CDKN1B gene in heart, kidney and lung of Tibetan pigs were extremely significantly or significantly higher than that of Yorkshire pigs (P<0.01 or P<0.05), and the expression of protein only in kidney tissue was significantly higher than that in Yorkshire pigs (P<0.05).【Conclusion】 The CDS sequence of CDKN1B gene in Tibetan pigs was successfully cloned.The expression of CDKN1B gene was high in heart, kidney and lung, and the expression in Tibetan pigs were higher than that in Yorkshire pigs.These results laid a foundation for further study on the role of CDKN1B gene and its encoded protein in hypoxia adaptation in Tibetan pigs.

Key words: Tibetan pigs; CDKN1B gene; bioinformatics; expression

CLC Number:

WEI Mingbang, BIANBA Qiongda, XIAO Qingqing, DUAN Mengqi, CHAMBA Yangzom, SHANG Peng. Cloning,Bioinformatics and Expression Analysis of CDKN1B Gene in Tibetan Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2196-2206.

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Cloning,Bioinformatics and Expression Analysis of CDKN1B Gene in Tibetan Pigs

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