EpCSP基因生物信息学分析及亚细胞定位

doi:10.19640/j.cnki.jtau.2025.04.003

天津农学院学报 ›› 2025, Vol. 32 ›› Issue (4): 18-25.doi: 10.19640/j.cnki.jtau.2025.04.003

EpCSP基因生物信息学分析及亚细胞定位

钟迎兰, 张富龙, 李欣雨, 杜锦, 黄海东, 吴疆^通信作者

天津农学院农学与资源环境学院,天津 300392

收稿日期:2024-01-08 发布日期:2025-09-02
通讯作者: 吴疆（1976—）,男,副教授,博士,主要从事植物分子生物学方面的研究。E-mail：wujiangjack@tjau.edu.cn。
作者简介:钟迎兰（1995—）,女,硕士在读,主要从事植物转基因方面的研究。E-mail：yinglanzhong@163.com。
基金资助:
天津市优秀企业科技特派员项目（22YDTPJC00960）

Bioinformatics analysis and subcellular localization of EpCSP gene

Zhong Yinglan, Zhang Fulong, Li Xinyu, Du Jin, Huang Haidong, Wu Jiang^{Corresponding Author}

College of Agronomy and Resource Environment, Tianjin Agricultural University, Tianjin 300392, China

Received:2024-01-08 Published:2025-09-02

摘要/Abstract

摘要： EpCSP基因是从微小杆菌（Exiguobacterium profundum）中克隆得到的一个冷激蛋白基因。生物信息学分析表明,该基因编码的蛋白可能定位于细胞核或细胞质基质。为进一步研究该基因功能,用PCR技术扩增EpCSP基因的CDS区,并将其克隆至表达载体pCAMBIA1300-EGFP-MCS上,构建用于瞬时表达EpCSP蛋白的重组质粒。经PCR鉴定后,重组质粒通过热激转化法转入农杆菌LBA4404,并用农杆菌介导法转化洋葱表皮细胞,用荧光显微镜观察确定其亚细胞定位。荧光显微镜观察结果显示,插入片段与预期序列完全一致,与载体构成了一个完整的基因表达系统,亚细胞定位结果表明EpCSP蛋白可能分布在细胞质基质。

关键词: EpCSP基因, 生物信息学, 洋葱表皮细胞, 亚细胞定位

Abstract: EpCSP gene is a cold shock protein gene cloned from Exiguobacterium profundum, and bioinformatics analysis suggests that the protein encoded by this gene may be located in the nucleus or cytoplasm matrix. To further investigate the function of the gene, the CDS region of the EpCSP gene was amplified using PCR technology and cloned onto the expression vector pCAMBIA1300-EGFP-MCS to construct a recombinant plasmid for transient expression of EpCSP protein. After PCR identification, the recombinant plasmid was transformed into Agrobacterium tumefaciens LBA4404 using electric shock transformation method and transformed into onion epidermal cells. Its subcellular localization was determined by fluorescence microscopy observation. The results showed that the inserted fragment was completely consistent with the expected sequence and formed a complete gene expression system with the vector. Subcellular localization results showed that the EpCSP protein was distributed in the cytoplasm matrix.

Key words: EpCsp gene, bioinformatics, onion epidermal cells, subcellular localization

中图分类号:

Q789

钟迎兰, 张富龙, 李欣雨, 杜锦, 黄海东, 吴疆. EpCSP基因生物信息学分析及亚细胞定位[J]. 天津农学院学报, 2025, 32(4): 18-25.

Zhong Yinglan, Zhang Fulong, Li Xinyu, Du Jin, Huang Haidong, Wu Jiang. Bioinformatics analysis and subcellular localization of EpCSP gene[J]. Journal of Tianjin Agricultural University, 2025, 32(4): 18-25.

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http://xuebao.tjau.edu.cn/CN/Y2025/V32/I4/18

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EpCSP基因生物信息学分析及亚细胞定位

Bioinformatics analysis and subcellular localization of EpCSP gene

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