多浪羊LIN28A基因启动子区克隆、序列分析及蛋白表达研究

doi:10.16431/j.cnki.1671-7236.2025.07.003

Abstract

Abstract: 【Objective】 The aim of this study was to explore the promoter sequence characteristics and protein expression of LIN28A gene in Duolang sheep,so as to lay a foundation for further study of its transcriptional regulation mechanism at puberty stage. 【Method】 Primers were designed for PCR amplification,cloning sequencing,and sequence alignment based on the LIN28A gene sequence of sheep in NCBI (accession No.:NC_056055.1),and the promoter sequence of LIN28A gene in Duolang sheep was obtained,which was analyzed by online analysis software.The hypothalamus,pituitary,and ovarian tissues of Duolang sheep during juvenile,prepuberty,puberty,and post-puberty periods were collected,and the expression of LIN28A protein in hypothalamic-pituitary-ovarian (HPO) axis was detected by Western blotting. 【Result】 The length of promoter sequence of LIN28A gene in Duolang sheep was 2 089 bp,and the GC content was 56.28%.The promoter sequence of LIN28A gene covered a total of 65 transcription factor binding sites,such as MEIS1,STAT3,and COMP1,3 TATA-boxes,1 CpG island,and 9 potential transcription start sites.LIN28A protein was expressed in all three tissues of Duolang sheep,with the highest expression in ovarian,followed by hypothalamus,and the lowest expression in pituitary.The expression of LIN28A protein in hypothalamus of Duolang sheep in prepuberty was significantly higher than that in juvenile and puberty (P＜0.05).The expression of LIN28A protein in pituitary of Duolang sheep in juvenile and prepuberty was significantly higher than that in puberty and post-puberty (P＜0.05).The expression of LIN28A protein in ovarian of Duolang sheep showed a stable trend before and after of puberty (P＞0.05).During the same period,there was no significant difference in the expression of LIN28A protein in hypothalamus,pituitary and ovarian of Duolang sheep (P＞0.05). 【Conclusion】 The promoter sequence of LIN28A gene in Duolang sheep had multiple transcription factor binding sites associated with puberty,providing experimental materials for transcription factor validation and promoter activity detection in the future.The expression of LIN28A protein in HPO axis of Duolang sheep was significantly different,providing a scientific basis for further study of the function and regulatory mechanism of LIN28A gene.

Key words: Duolang sheep; LIN28A gene; promoter; puberty; HPO axis

CLC Number:

S826.2

LI Wei, HUANG Qiaoyan, WANG Xinkun, GU Ruohuai, SUN Huiping, ZHU Lexiao, XING Feng. Cloning, Sequence Analysis and Protein Expression of LIN28A Gene Promoter in Duolang Sheep[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 2992-3003.

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Cloning, Sequence Analysis and Protein Expression of LIN28A Gene Promoter in Duolang Sheep

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