首页|水稻OsEnS51可变剪接及编码蛋白结构分析

水稻OsEnS51可变剪接及编码蛋白结构分析

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本研究综合运用生物信息学、RT-PCR和TA克隆技术对水稻胚乳特异表达基因OsEnS51的可变剪接本进行鉴定,并分析剪接本的表达模式和蛋白质结构.利用生物信息学分析发现,该基因可能存在4种可变剪接转录本.结构域分析发现,OsEnS51.1和OsEnS51.2含有2个Cupin_1结构域,而OsEnS51.3和OsEnS51.4仅含有1个Cupin_1结构域,表明OsEnS51不同可变剪接本所表达的蛋白存在结构分化.根据不同可变剪接本的序列信息设计特异性引物,进行RT-PCR扩增,结合TA克隆和测序成功鉴定到其中3种可变剪接转录本,分别是OsEnS51.1、OsEnS51.3和OsEnS51.4.表达分析结果显示OsEnS51.1~OsEnS51.4在授粉后7~15d的种子表达较高,且随着种子的发育其表达不断增加;另外OsEnS51.4的表达量比OsEnS51.1高,表明OsEnS51不同可变剪接本的含量存在差异.综上,OsEnS51至少存在3种不同可变剪接本,且编码的蛋白存在结构分化,本研究结果为进一步研究OsEnS51的生物学功能提供了基础.
Alternative Splicing of Rice OsEnS51 and Structural Analysis of Encoding Protein
In this study,bioinformatics,RT-PCR and TA cloning techniques were used to identify alternatively spliced transcripts of rice endosperm-specific expression gene OsEnS51,and the expression of the spliceosome were then analyzed.4 potential alternatively spliced transcripts were predicted with bioinformatics analysis.Of these tran-scripts,OsEnS51.1 and OsEnS51.2 contains 2 Cupin_1 domains,while OsEnS51.3 and OsEnS51.4 only contains 1 Cupin_1 domain respectively,suggesting structural differentiation of various OsEnS51 transcripts.According to the sequence information of different alternatively spliced transcripts,specific primers were designed for further RT-PCR amplification.Combined with TA cloning and sequencing,three of them were successfully identified alterna-tively spliced transcripts,designated OsEnS51.1,OsEnS51.3 and OsEnS51.4.OsEnS51.1~OsEnS51.4 was high ex-pressed in seeds 7~15 days following pollination,whose transcription products continued to accumulate with the de-velopment of seeds.Higher expression level of OsEnS51.4 was detected compared to that of OsEnS51.1,indicating divergent expression patterns of different alternatively spliced transcripts of OsEnS51.Altogether,OsEnS51 has at least 3 alternative splicing transcripts with differentiated encoding protein structures.These results laid the founda-tion for further research on the biological functions of OsEnS51.

Oryza sativaSeed-specifically expressed genesAlternative splicingExpression analysisThree-dimensional protein structure

符霖、王慧慧、黄思琳、阎新、王鑫

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南昌大学生命科学学院,江西省分子生物学与基因工程重点实验室,南昌,330031

水稻 种子特异表达基因 可变剪接 表达分析 蛋白质三维结构

国家自然科学基金项目江西省研究生创新专项资金项目

32060135YC2019-B021

2024

分子植物育种
海南省生物工程协会

分子植物育种

CSTPCD北大核心
影响因子:0.765
ISSN:1672-416X
年,卷(期):2024.22(19)