槟榔江水牛<i>SREBF1</i>基因克隆、分子特征及组织表达分析

刀文彬; 欧阳依娜; 梁建平; 范新阳; 陈馨瑶; 苗永旺

doi:10.12101/j.issn.1004-390X(n).202302037

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槟榔江水牛SREBF1基因克隆、分子特征及组织表达分析

刀文彬^{1, 2, #,},
欧阳依娜^{1, 2, 3, #,},
梁建平⁴,
范新阳^{1, 2},
陈馨瑶⁵,
苗永旺^{1, 2, ,}

1.
云南农业大学动物科学技术学院，云南昆明 650201
2.
云南农业大学动物遗传育种研究所，云南昆明 650201
3.
云南省畜牧兽医科学院，云南昆明 650224
4.
云南省德宏州科学技术创新中心，云南芒市 678400
5.
腾冲市巴福乐槟榔江水牛良种繁育有限公司，云南腾冲 679118

基金项目: 国家自然科学基金项目(32260822，31760659)；云南省重大科技专项计划项目(202202AE090005)。

详细信息

作者简介:
#对本文贡献等同，为并列第一作者。刀文彬(1997—)，男，云南普洱人，在读硕士研究生，主要从事动物分子遗传学研究。E-mail：dwbin3@163.com

欧阳依娜(1979—)，女，云南昆明人，博士，主要从事动物遗传学研究。E-mail：yinaouyang@126.com

通信作者:
苗永旺(1964—)，男，内蒙古通辽人，博士，教授，主要从事动物遗传学研究。E-mail：yongwangmiao1@126.com

#对本文贡献等同，为并列第一作者。

中图分类号: S823.912
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出版历程
- 收稿日期: 2023-02-14
- 修回日期: 2025-02-05
- 录用日期: 2025-02-15
- 网络出版日期: 2025-02-18

摘要:

目的

阐明水牛固醇调节元件结合转录因子1 (sterol regulatory element binding transcription factor 1，SREBF1)的分子特征和组织表达特异性，初步揭示其分子功能。

方法

以槟榔江水牛为研究对象，采用RT-PCR技术克隆水牛SREBF1基因完整编码序列(coding sequence，CDS)；利用生物信息学方法分析SREBF1分子特征；采用qPCR技术检测SREBF1在泌乳期水牛多个组织中的表达特异性。

结果

水牛SREBF1 CDS全长3438 bp，编码1145个氨基酸残基组成的亲水蛋白；SREBF1包含1个bHLHzip_SREBP1 (AA319~393) DNA结合结构域，无跨膜结构和信号肽，主要定位于细胞核和内质网，主要功能是结合并调控DNA转录、参与脂质代谢等。水牛与其他牛科物种SREBF1在一级结构、理化特征、结构域和高级结构上高度相似。在基于SREBF1氨基酸序列构建的系统发育树上，水牛与其他牛科物种聚为1类。在泌乳期水牛的8个组织中，SREBF1基因在乳腺中的表达水平最高，其次是肝脏、骨骼肌、垂体、肾脏，在脾脏、瘤胃和心脏中的表达水平较低。

结论

水牛与其他牛科物种SREBF1氨基酸序列具有高度的相似性。水牛SREBF1可调控DNA转录，在脂质代谢旺盛的组织中发挥作用。

关键词:

Cloning, Molecular Characteristics and Tissue Expression Analysis of SREBF1 Gene in Binglangjiang Buffalo

DAO Wenbin^{1, 2, #,},
OUYANG Yina^{1, 2, 3, #,},
LIANG Jianping⁴,
FAN Xinyang^{1, 2},
CHEN Xinyao⁵,
MIAO Yongwang^{1, 2, ,}

1.
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China
2.
Institute of Animal Genetics and Breeding, Yunnan Agricultural University, Kunming 650201, China
3.
Yunnan Animal Science and Veterinary Institute, Kunming 650224, China
4.
Dehong Science and Technology Innovation Center of Yunnan Province, Mangshi 678400, China
5.
Tengchong Co., Ltd. of Fine Variety Breeding of Binglangjiang Buffalo, Tengchong 679118, China

Abstract:

Purpose

To elucidate the molecular characteristics and tissue expression specificity of buffalo sterol regulatory element binding transcription factor 1 (SREBF1), revealing the molecular function preliminarily.

Methods

The Binglangjiang buffalo was used as research object, and the complete coding sequence (CDS) of buffalo SREBF1 gene was cloned by RT-PCR. The molecular characteristics of SREBF1 were analyzed by bioinformatics method. The expression specificity of SREBF1 gene in multiple tissues of lactating buffalo was further detected by qPCR.

Results

The CDS length of buffalo SREBF1 gene was 3 438 bp, it encoded a protein composed of 1 145 amino acids. The protein was hydrophilic and contained a bHLHzip_SREBP1 (AA319-393) domain, and without transmembrane structure and signal peptide. The SREBF1 protein was localized in the nucleus and endoplasmic reticulum. The main functions were to bind and regulate DNA transcription, and participate in lipid metabolism. The primary structure, physicochemical characteristics, the composition of domains, and advanced structure of SREBF1 protein were highly similar among buffalo and other Bovidae species. Phylogenetic analysis of amino acid sequence based on SREBF1 showed buffalo clustered with other Bovidae species. The expression in eight tissues of lactating buffaloes showed that SREBF1 gene expression was higher in mammary gland, followed by liver, skeletal muscle, pituitary gland and kidney, and lower in spleen, rumen and heart.

Conclusion

The amino acid sequence of SREBF1 in buffalo and other Bovidae species is highly similar. Buffalo SREBF1 can regulate DNA transcription and play a role in tissues with high lipid metabolism.

Keywords:

参考文献

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