无量山乌骨鸡HMOX1基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2023.11.007

Abstract

Abstract: 【Objective】 The purpose of this study was to amplify the CDS region of heme oxygenase 1 (HMOX1) gene in Wuliangshan Sooty chickens and carry out the bioinformatics analysis, and detect the tissue expression characteristics of HMOX1 gene, so as to lay a foundation for the subsequent functional research of HMOX1 gene.【Method】 The HMOX1 gene was amplified by PCR using the adipose tissue cDNA in Wuliangshan Sooty chickens as the template, and the PESI-T vector was connected for sequencing analysis.The obtained sequence was translated by DNAMAN software, and the sequence was compared with that of Gallus gallus.BLAST, Mega 7.0, SOPMA, ProtParam and ExPASy were used to carry out the similarity alignment, phylogenetic tree construction and the prediction of physicochemical properties and protein structure.STRING software was used to predict HMOX1 interacting proteins.The expression of HMOX1 gene in different tissues of Wuliangshan Sooty chickens were detected by Real-time quantitative PCR.【Result】 The length of CDS region of HMOX1 gene in Wuliangshan Sooty chickens was 891 bp, encoding 296 amino acids.Three mutations were found in the nucleotide of HMOX1 gene with that of Gallus gallus (accession No.:NM_205344.1), two synonymous mutations, G>C at 217 bp was a missense mutation, resulting in aspartic acid mutation at 73 to glycine.The higher amino acid similarity of HMOX1 gene were 99.6% and 95.0% with Gallus gallus and Coturnix japonica, respectively, and the lowest was 60.2% with Equus caballus.Phylogenetic tree analysis showed that Wuliangshan Sooty chickens had the closer genetic relationship with Gallus gallus and Coturnix japonica, and was far away from non poultry. HMOX1 protein was unstable and hydrophilic, was mainly located in cytoplasm, and there was no signal peptide cleavage site, it was a non secreted protein.There were 1 transmembrane helix structure and 47 phosphorylation sites.The secondary structure of HMOX1 protein was mainly alpha helix.HMOX1 protein might interact with 10 proteins such as HMOX2, BLVRA, HEPH, CP and COX10.The tissue expression analysis showed that HMOX1 gene was expressed in all tissues of Wuliangshan Sooty chickens, with the highest expression in liver and the lowest expression in heart.【Conclusion】 The CDS length of HMOX1 gene in Wuliangshan Sooty chickens was 891 bp, encoding 296 amino acids, predicting it interacted with 10 proteins.HMOX1 gene was highly expressed in liver.The results provided a theoretical basis of informatics for further exploring the role of HMOX1 gene in chicken fat deposition.

Key words: Wuliangshan Sooty chickens; HMOX1 gene; cloning; bioinformatics; tissue expression

CLC Number:

S831.2

OU Xiaoman, LIU Minghuan, WU Yun, GUO Aiwei, LI Qiuyu, CHEN Fenfen. Cloning, Bioinformatics and Tissue Expression Analysis of HMOX1 Gene in Wuliangshan Sooty Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4381-4391.

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Cloning, Bioinformatics and Tissue Expression Analysis of HMOX1 Gene in Wuliangshan Sooty Chickens

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