雪山草鸡血管紧张素转换酶2(ACE2)基因克隆、结构特性分析及其表达特征研究

doi:10.11843/j.issn.0366-6964.2021.011.007

Abstract

Abstract: The study aimed to clone the angiotensin converting enzyme 2 (ACE2) gene, analyze its structural and expressing characteristics. In this experiment, the Xueshan chicken was selected as the research objects, the coding region of ACE2 gene was amplified by PCR, the structural characteristics were analyzed by the method of bioinformatics, the standard plasmid of ACE2 gene was constructed by TA cloning, and the expression characteristics of ACE2 gene was determined in the tissues of Xueshan chicken by absolute quantitative method. The results showed that CDS (2 427 bp) of ACE2 was cloned and encoded 808 amino acids, it was consistent with the ACE2 gene in white-feathered broilers, but a large number of mutations were found in yellow-feathered broilers which led to changes in amino acids. The homology analysis found that sequence of ACE2 gene of Xueshan chicken had 99% homology with red jungle fowl, followed by mallard duck 88%, and 73%-76% with other animals. The bioinformatics analysis showed that ACE2 belonged to type I transmembrane protein(secretory protein), and signal peptide was located in 1-17aa, contained 46 protein phosphorylation sites. The quantitative results found that the expression of ACE2 in the small intestine was significantly higher than that in other tissues. The expression of ACE2 in bursa, trachea, gallbladder and spleen was lower, and the expression in the spleen was the lowest. The expression of ACE2 in jejunum, cecum, spleen and lung of hens were significantly higher than those of roosters. The expression of ACE2 in kidney and heart of roosters were significantly higher than those of hens. The expression of ACE2 in the duodenum, jejunum, lung, spleen and ileum tissues of 90-day-old Xueshan chicken were significantly lower than those of 45-day-old Xueshan chicken, and the results showed an opposite trend in the bursa and kidney. The coding region of ACE2 gene was obtained in Xueshan chicken. Bioinformatics analysis showed that ACE2 gene was conservative in poultry. The expression level of ACE2 gene was higher in the intestinal tissues than that in other tissues, higher in hen than that in rooster, and the expression level decreased with time. The results laid a foundation for the study of function of ACE2 gene in the gut of yellow feather broilers.

Key words: Xueshan chicken, angiotensin converting enzyme 2 gene, sequence analysis, tissues expression characteristics

CLC Number:

S831.2

CHEN Qingyi, CONG Guanglei, XIAO Yunqi, SHI Shourong. Molecular Cloning, Sequence Analysis and Expression Characteristics of Angiotensin Converting Enzyme 2 (ACE2) Gene in Xueshan Chicken[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3053-3063.

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Molecular Cloning, Sequence Analysis and Expression Characteristics of Angiotensin Converting Enzyme 2 (ACE2) Gene in Xueshan Chicken

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