绵羊CREBRF基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2022.05.001

Abstract

Abstract: 【Objective】 The purpose of this study was to clone Luman/CREB3 recruitment factor(CREBRF) gene of Ovis aries and analyze its bioinformatics,and detect the expression of CREBRF gene in different tissues of Ovis aries,so as to provide theoretical reference for exploring the biological function of CREBRF gene in Ovis aries.【Methods】 Using ovary cDNA of Ovis aries as template,the complete CDS region of CREBRF gene was amplified by PCR and cloned,the similarity alignment,phylogenetic tree and bioinformatics analysis were carried out.The expression of CREBRF gene in different tissues of Ovis aries was detected by Real-time quantitative PCR.【Result】 The total length of CDS region of CREBRF gene was 1 920 bp,encoding 639 amino acids.The similarity alignment results showed that the similarity of CREBRF amino acid sequence of Ovis aries with Capra hircus,Bos taurus,Homo sapiens,Mus musculus,Sus scrofas,Canis lupus familiaris,Equus przewalskii,Gallus gallus,Anas platyrhynchos and Danio rerio were 99.8%,99.1%,95.4%,93.6%,98.3%,97.5%,98.4%,88.0%,87.5% and 61.3%,respectively.The phylogenetic tree analysis showed that Ovis aries had the closest relationship with Capra hircus and Bos taurus,and the furthest relationship with Danio rerio.Bioinformatics analysis found that the molecular formula of CREBRF protein in Ovis aries was C₃₁₂₆H₄₉₁₄N₈₅₈O₁₀₅₆S₂₁,the theoretical molecular weight was 72.08 ku,the theoretical isoelectric point (pI) was 4.77,the half-life was 30 h,the N-terminal of peptide chain was methionine (Met),and the instability coefficient was 54.83.CREBRF protein existed in the nucleus,there was no transmembrane property and signal peptide,and was a hydrophilic unstable protein.The secondary structure of CREBRF protein was mainly random random coil (47.57%),followed by alpha helix(37.09%).Real-time quantitative PCR results showed that CREBRF gene was expressed in different tissues of Ovis aries,the expression in heart,kidney and ovary were significantly higher than that in other tissues (P<0.05).【Conclusion】 This experiment obtained the full-length sequence of CREBRF gene,and preliminarily studied its tissue expression,which provided materials for studying the regulatory mechanism of embryonic development and improving fecundity in Ovis aries.

Key words: Ovis aries; CREBRF gene; bioinformatics analysis; expression

CLC Number:

ZHANG Limeng, LIU Aiju, LI Runting, LI Yuhua, LI Lin, NIE Xiaoning, WANG Linqing, CHEN Longxin. Cloning,Bioinformatics and Tissue Expression Analysis of CREBRF Gene in Ovis aries[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1599-1609.

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Cloning,Bioinformatics and Tissue Expression Analysis of CREBRF Gene in Ovis aries

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