水稻OsEnS51可变剪接及编码蛋白结构分析  

Alternative Splicing of Rice OsEnS51 and Structural Analysis of Encoding Protein

作　　者：符霖 王慧慧 黄思琳 阎新[1] 王鑫 Fu Lin;Wang Huihui;Huang Silin;Yan Xin;Wang Xin(Key Laboratory of Molecular Biology and Genetic Engineering of Jiangxi Province,School of Life Sciences,Nanchang University,Nanchang,330031)

机构地区：[1]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

出　　处：《分子植物育种》2024年第19期6265-6271,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(32060135);江西省研究生创新专项资金项目(YC2019-B021)共同资助。

摘　　要：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7~15 d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。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 transcripts,OsEnS51.1 and OsEnS51.2 contains 2 Cupin_1 domains,while OsEnS51.3 and OsEnS51.4 only contains 1Cupin_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 RTPCR amplification.Combined with TA cloning and sequencing,three of them were successfully identified alternatively spliced transcripts,designated OsEnS51.1,OsEnS51.3 and OsEnS51.4.OsEnS51.1~OsEnS51.4 was high expressed in seeds 7~15 days following pollination,whose transcription products continued to accumulate with the development 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 foundation for further research on the biological functions of OsEnS51.

关 键 词：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 

分 类 号：S511[农业科学—作物学]