番茄MYB44基因生物信息学及亚细胞定位分析

doi:10.11924/j.issn.1000-6850.casb2020-0807

摘要/Abstract

摘要：

旨在分析SlMYB44基因的生物学功能,为其抗冷害作用机制提供科学依据。本文以俄罗斯栽培番茄Bolgogragsky为材料,根据转录组测序结果,筛选出差异表达基因SlMYB44作为研究对象。运用多种生物信息学数据库分析了番茄SlMYB44基因的保守结构域、亲疏水性、信号肽以及启动子顺式作用元件。并且构建了pMDC43-MYB44表达载体进行亚细胞定位分析。结果表明SlMYB44基因全长1551 bp,共372个氨基酸。SlMYB44蛋白没有潜在的信号肽和跨膜结构,属于不稳定、亲水性蛋白。SlMYB44基因启动子顺式作用元件分析发现,大部分与逆境胁迫、激素响应等相关。亚细胞定位分析发现SlMYB44基因表达产物定位于细胞核中。SlMYB44基因对番茄冷害有响应,进一步推测其可能影响番茄御冷机制。

关键词: 番茄, 基因, SlMYB44, 生物信息学分析, 亚细胞定位, 低温胁迫

Abstract:

The aims are to analyze the biological function of SlMYB44 gene and provide a scientific basis for exploring its mechanism of resisting chilling injury. In this study, the Russian cultivated tomato bolgogragsky was used as the material, and according to the sequencing results of transcriptome, the differentially expressed gene SlMYB44 was selected as the research object. The conservative functional domain, hydrophilicity or hydrophobicity, signal peptide and promoter cis-acting elements of tomato SlMYB44 gene were analyzed by using various bioinformatics databases. The expression vector pMDC43-MYB44 was constructed for subcellular localization analysis. SlMYB44 gene encoded 372 amino acids with a total length of 1551 bp. SlMYB44 protein had no potential signal peptide site and membrane spaning domain, which belonged to labile protein and hydrophilic protein. The analysis of cis-acting elements of SlMYB44 gene promoter showed that most of them were related to adversity stress and hormone response. Subcellular localization analysis demonstrated that the expression product of SlMYB44 gene was localized in the cell nucleus. In conclusion, the SlMYB44 gene responds to chilling injury, it is further speculated that it might affect the tomato chilling mechanism.

Key words: Solanum lycopersicum, gene, SlMYB44, bioinformatics analysis, subcellular localization, low temperature stress

中图分类号:

S641.2

李颖, 王莹莹, 苏旭, 谷建田. 番茄MYB44基因生物信息学及亚细胞定位分析[J]. 中国农学通报, 2021, 37(14): 34-40.

Li Ying, Wang Yingying, Su Xu, Gu Jiantian. MYB44 Gene from Solanum Lycopersicum: Bioinformatics Analysis and Subcellular Localization[J]. Chinese Agricultural Science Bulletin, 2021, 37(14): 34-40.

图/表 11

参考文献 21

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类型	名称	序列	数量	物种
参与干旱诱导	MBS	CAACTG	6	Arabidopsis thaliana
参与厌氧反应	ARE	AAACCA	6	Zea mays
参与厌氧反应	O₂-site	GATGA(C/T)(A/G)TG(A/G)	8	Zea mays
参与光反应	Box 4	ATTAAT	6	Petroselinum crispum
	ATCT-motif	AATCTAATCC	9	Pisum sativum
	TCT-motif	TCTTAC	18	Arabidopsis thaliana
	I-box	GTATAAGGCC	9	Larix laricina
	GATA-motif	AAGGATAAGG	9	Solanum tuberosum
启动子和增强子区域	CAAT-box	CCAAT	30	Arabidopsis thaliana
启动子和增强子区域	CAAT-box	CAAAT	20	Pisum sativum
参与MeJA反应	TGACG-motif	TGACG	15	Hordeum vulgare
参与MeJA反应	CGTCA-motif	CGTCA	15	Hordeum vulgare
参与赤霉素反应	TATC-box	TATCCCA	7	Oryza sativa
核心启动子元素转录	TATA-box	TATA	16	Arabidopsis thaliana
	TATA-box	ATATAA	12	Brassica napus
	TATA-box	TATAA	5	Arabidopsis thaliana

类型	名称	序列	数量	物种
参与干旱诱导	MBS	CAACTG	6	Arabidopsis thaliana
参与厌氧反应	ARE	AAACCA	6	Zea mays
参与厌氧反应	O₂-site	GATGA(C/T)(A/G)TG(A/G)	8	Zea mays
参与光反应	Box 4	ATTAAT	6	Petroselinum crispum
	ATCT-motif	AATCTAATCC	9	Pisum sativum
	TCT-motif	TCTTAC	18	Arabidopsis thaliana
	I-box	GTATAAGGCC	9	Larix laricina
	GATA-motif	AAGGATAAGG	9	Solanum tuberosum
启动子和增强子区域	CAAT-box	CCAAT	30	Arabidopsis thaliana
启动子和增强子区域	CAAT-box	CAAAT	20	Pisum sativum
参与MeJA反应	TGACG-motif	TGACG	15	Hordeum vulgare
参与MeJA反应	CGTCA-motif	CGTCA	15	Hordeum vulgare
参与赤霉素反应	TATC-box	TATCCCA	7	Oryza sativa
核心启动子元素转录	TATA-box	TATA	16	Arabidopsis thaliana
	TATA-box	ATATAA	12	Brassica napus
	TATA-box	TATAA	5	Arabidopsis thaliana

引物ID	引物序列(5'-3')	注释
L2162	AtgaactatacaaaggcgcgccaATGGCGATTACTACAAACAG	pMDC43-MYB44带有AscI酶切位点的正向引物
L2163	GatcggggaaattcgagctcCTAATCGAATTTGCTTAGTC	pMDC43-MYB44带有 SacI酶切位点的反向引物
43F	ccacacaatctgccctttcg	pMDC43正向引物
43R	cggcaacaggattcaatcttaag	pMDC43反向引物

引物ID	引物序列(5'-3')	注释
L2162	AtgaactatacaaaggcgcgccaATGGCGATTACTACAAACAG	pMDC43-MYB44带有AscI酶切位点的正向引物
L2163	GatcggggaaattcgagctcCTAATCGAATTTGCTTAGTC	pMDC43-MYB44带有 SacI酶切位点的反向引物
43F	ccacacaatctgccctttcg	pMDC43正向引物
43R	cggcaacaggattcaatcttaag	pMDC43反向引物