赤水乌骨鸡PITX1基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2023.10.003

Abstract

Abstract: 【Objective】 The purpose of this study was to clone the paired-like homeodomain transcription factor 1 (PITX1) gene in Chishui Black bone chickens and carry out bioinformatics analysis,and detect the expression of PITX1 gene in different tissues of Chishui Black bone chickens,so as to provide the materials for biological function of PITX1 gene in muscle growth and development of chickens.【Method】 The sequence of PITX1 gene CDS in Chishui Black bone chickens was cloned and sequenced using breast muscle cDNA as template.The alignment and bioinformatics analysis were carried out by online softwares.The expressions of PITX1 gene in heart,liver,spleen,kidney,lung,breast and leg muscles in Chishui Black bone chickens were detected by Real-time quantitative PCR.【Result】 The full length sequence of PITX1 gene CDS in Chishui Black bone chickens was 909 bp,which had 5 synonymous mutations,and encoding 302 amino acids.The amino acid sequence similarity of PITX1 gene in Chishui Black bone chickens were 100%,99.7%,99.0%,85.0% and 84.4% with Gallus gallus,Anas platyrhynchos,Coturnix japonica, Oryctolagus cuniculus and Mus musculus,respectively,and the sequence similarity with Homo sapiens,Bos taurus and Sus scrofa all were 85.7%.The results of phylogenetic tree analysis showed that Chishui Black bone chickens was the closest to Gallus gallus and the furthest to Homo sapiens.PITX1 protein of Chishui Black bone chickens was a hydrophilic protein,which had 47 phosphorylation sites,1 N-glycosylation modification site and 15 O-glycosylation modification sites,without transmembrane domain and signal peptide,and mainly distributed in nucleus.The secondary structure was composed of random coil,alpha helix,extended chain and beta turn,the tertiary structure prediction was consistent with it.Protein interaction prediction analysis indicated that PITX1 might be interacted with BARX1,NKX2-8,POU1F1,EGR1,NR5A1,CTNNB1,TBX5 and TBX4.Real-time quantitative PCR results revealed that PITX1 gene was expressed in different tissues of Chishui Black bone chickens,and the highest expression was found in lung,followed were liver and leg muscle,but that was the lowest in breast muscle.【Conclusion】 The sequence of PITX1 gene CDS in Chishui Black bone chickens was 909 bp,and encoding 302 amino acids,that was a hydrophilic protein,and mainly distributed in nucleus.PITX1 gene was expressed in all tissues of Chishui Black bone chickens,which was the highest in lung.The results provided a reference for further study on the function of PITX1 gene in chickens.

Key words: Chishui Black bone chickens; PITX1 gene; bioinformatics; expression

CLC Number:

WU Yun, OU Zhengmiao, LI Qiuyu, CHEN Fenfen, YAN Jimei. Cloning,Bioinformatics and Tissue Expression Analysis of PITX1 Gene in Chishui Black Bone Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 3896-3905.

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Cloning,Bioinformatics and Tissue Expression Analysis of PITX1 Gene in Chishui Black Bone Chickens

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