半番鸭NMB基因克隆、生物信息学和组织表达分析

doi:10.16431/j.cnki.1671-7236.2025.04.023

Abstract

Abstract: 【Objective】 The purpose of this study was to clone and sequence the neuromedin B (NMB) gene from Mulard duck,and to detect the expression of NMB gene in Mulard duck.【Method】 According to the predicted NMB gene sequence of duck in the NCBI database (accession No.：XM_027466185.2),primers were designed to amplify NMB gene fragment in Mulard duck by PCR amplification,and the NMB gene sequence was obtained by cloning and sequencing.The nucleotide and amino acid sequences of NMB in Mulard duck were analyzed using bioinformation analysis software,and the phylogenetic tree was constructed.Real-time quantitative PCR was used to detect the expression of NMB gene mRNA in multiple tissues of Mulard duck.【Result】 In this study,NMB gene of Mulard duck was cloned with 387 bp CDS sequence and encoding 121 amino acids,including 14 amino acids (GNLWATGHFMGKKS) of the conserved domain of bombesin,which was identical to that in NMB of Gallus gallus，Mus musculus，Sus crofa，Bos taurus and Homo sapiens.The similarity analysis showed that the similarity of nucleotide and amino acid sequences of NMB between Mulard duck and Gallus gallus were 87.9% and 89.8%,respectively,and the similarity with other animals were lower.In addition,NMB of Mulard duck was evolutionarily close to Gallus gallus,and far from domestic animals such as Bos taurus, Equus caballus,Sus crofa and Canis lupus familiaris.The results of bioinformatics analysis showed that NMB protein of Mulard duck was a hydrophilic protein,and its N-terminal 22 amino acids were signal peptides.In the secondary and tertiary structures,the proportion of alpha helix,extended chain,beta turn and random coil were 53.91%,14.06%,7.03% and 25.00%,respectively.The NMB protein of Mulard duck mainly binded to NMBR and GRPR proteins.Real-time quantitative PCR results showed that NMB gene mRNA was expressed in the central and peripheral tissues and organs of Mulard duck.Compared with the jejunum,the expression levels of NMB gene were relatively high in the central cerebellum,brain and optic lobe (P＜0.01),and relatively high in the peripheral trachea,lung,pineal gland,thyroid,kidney and uropygial gland (P＜0.01).In addition,the expression trend of NMB gene mRNA in Mulard duck of different genders was basically the same,but there were different expression in female and male ducks in individual tissues.The expression of NMB gene mRNA in female duck was relatively higher than that in male duck in cerebellum and pancreas (P＜0.01),but the opposite was true in pineal gland and kidney (P＜0.01 or P＜0.05).【Conclusion】 NMB protein in Mulard duck had the conserved domain of bombesin.The genetic relationship was the closest between Mulard duck and Gallus gallus,and the protein similarity was high.NMB protein was hydrophilic protein and had signal peptide.NMB gene was highly expressed in the central (cerebellum,brain and optic lobe) and peripheral (pineal gland,kidney,uropygial gland,thyroid gland and trachea,etc.) tissues and organs of Mulard duck.The results could provide reference for further study on the function of NMB gene in Mulard duck.

Key words: Mulard duck; NMB gene; cloning; expression; bioinformatics

CLC Number:

S834⁺.89

ZHANG Ying, CHEN Guangzhen, LIU Mei, LI Hongxia, YU Yan, JIA Xuebo, LI Wenqian, FAN Jianhua, MA Zhiyu. Cloning,Bioinformatics and Tissue Expression Analysis of NMB Gene in Mulard Duck[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(4): 1705-1715.

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Cloning,Bioinformatics and Tissue Expression Analysis of NMB Gene in Mulard Duck

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