多浪羊NPTX1基因克隆、生物信息学分析及在初情期的表达研究

doi:10.16431/j.cnki.1671-7236.2023.03.020

Abstract

Abstract: 【Objective】 This study was aimed to clone the coding region (CDS) sequence of neuronal pentraxin 1 (NPTX1) gene in Dolang sheep,analyze its structure and function by bioinformatics,and study its expression pattern in different tissues of gonadal axis during puberty initiation in Dolang sheep.【Method】 The hypothalamus,pituitary,oviduct,ovary and uterus tissues of healthy female Dolang sheep were collected from prepubertal,pubertal and postpubertal.The total RNA was extracted and reverse transcribed into cDNA.NPTX1 gene of Dolang sheep was amplified by PCR using prepubertal hypothalamus cDNA as template,cloned and sequenced,and its bioinformatics analysis was carried out.The expression of NPTX1 gene in hypothalamus,pituitary,ovary,uterus and oviduct of Dolang sheep were analyzed by Real-time quantitative PCR.【Result】 The cloned NPTX1 gene was 1 576 bp,of which the CDS sequence was 1 299 bp,and the similarity with the mRNA sequence of sheep predicted on GenBank(accession No.:XM_005683183.3) was 99.85%.Phylogenetic tree analysis results showed that NPTX1 gene of Dolang sheep had the closest genetic distance to Ovis aries and the farthest genetic distance to Gallus gallus.Bioinformatics analysis results showed that NPTX1 protein was an unstable hydrophilic acidic protein with a signal peptide and belonged to secretory protein.In the secondary structure of NPTX1 protein,alpha helix and random coil accounted for 41.90% and 40.73%,respectively,which were consistent with the predicted results of the tertiary structure.Real-time quantitative PCR results showed that NPTX1 gene was expressed in five different gonad tissues of Dolang sheep in three stages,and the expression was the highest in hypothalamus of puberty,which was significantly higher than that of prepuberty and postpuberty (P＜0.05).【Conclusion】 The experiment successfully cloned the full-length CDS sequence of NPTX1 gene in Dolang sheep,and revealed the differences in the expression of NPTX1 gene in gonadal axis tissue of Dolang sheep at different stages of puberty,which provided a basis for further exploring the regulation mechanism of NPTX1 gene in puberty onset of Dolang sheep.

Key words: Dolang sheep; NPTX1 gene; cloning; puberty; tissue expression

CLC Number:

S813.3

ZHANG Yongjie, SUI Zhiyuan, ZHANG Zhishuai, WANG Chenguang, LI Xiaojun, XING Feng. Cloning and Bioinformatics Analysis of NPTX1 Gene and Its Tissues Expression at Puberty in Dolang Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1048-1058.

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Cloning and Bioinformatics Analysis of NPTX1 Gene and Its Tissues Expression at Puberty in Dolang Sheep

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