水稻OsJMJ718基因可选择性多聚腺苷酸化序列的克隆及生殖发育期表达模式  

Cloning and Expression Pattern Analysis of Rice OsJMJ718 Alternative Polyadenylation Sequences During Reproductive Developmental Stage

作　　者：鲁丹[1] 王丽[1] 宋凡[1] 陶菊红 张大兵[1] 袁政[1] Dan Lu;Li Wang;Fan Song;Juhong Tao;Dabing Zhang;Zheng Yuan＊(School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学生命科学技术学院,上海200240

出　　处：《植物学报》2018年第5期594-602,共9页Chinese Bulletin of Botany

基　　金：国家七大农作物育种专项(No.2016YFD0101107);上海市科委基础研究重大重点项目(No.14JC1403901);国家自然科学基金(No.31470397;No.31671260)

摘　　要：可选择性多聚腺苷酸化是真核生物重要的基因调控机制之一,通过形成不同长度的3'端非翻译区影响信使RNA的稳定性、定位和翻译效率,从而增加转录本的复杂度。已有研究表明,拟南芥(Arabidopsis thaliana)中参与DNA去甲基化调控IBM1基因的可选择性多聚腺苷酸化加工受染色质调节因子EDM2调控,从而影响拟南芥基因组数千基因的CHG甲基化水平,但该类调控机制是否在其它物种中同样存在仍然未知。以水稻(Oryza sativa)基因组中IBM1同源基因Os JMJ718为研究对象,利用生物信息学分析和3'RACE实验,发现IBM1同源基因也存在可选择性多聚腺苷酸化修饰,Os JMJ718基因可能存在9个可选择性多聚腺苷酸化序列。序列比对分析表明,NCBI网站现存日本晴Os JMJ718基因组3'末端序列与9522和明辉63等其它生态型基因组序列组成可能不同。荧光实时定量PCR分析表明,Os JMJ718的9个转录本在水稻生殖发育阶段呈现不同的动态表达模式,其中TVX5转录本表达量最高。研究获得的Os JMJ718基因可选择性多聚腺苷酸化序列信息及相关的表达模式分析为进一步揭示水稻Os JMJ718基因的可选择性多聚腺苷酸化分子机制和生物学功能奠定了基础。Alternative polyadenylation （APA） is one of the important regulatory pathways of eukaryotic gene expression. By forming different lengths of 3＇ untranslated regions, APA affects the stability, localization and translation efficiency of mRNA and increases the complexity of transcripts. The expression of the Arabidopsis gene increased polyadenylation of BONSAI methylation I （IBM1） is regutated by chromatin regulatory factor enhanced downy mildew 2 （EDM2）, which can further affect the CHG methylation level of the Arabidopsis thafiana genome. However, whether such regulatory mecha- nisms exist in other species is unknown. To answer this question, we selected OsJMJ718, an IBM1 homologous gene of rice, for research. By using bioinformatics analysis and 3＇RACE experiments, we found that homologous genes of IBM1 also had APA modification. Among them, the OsJMJ718 gene may have 9 alternative polyadenylation sequences. Further sequence alignment analysis revealed that the 3＇ terminal sequence of OsJMJ718 in the existing Japonica genome in the NCBI database may be different from that of the other ecotype genome sequences, such as 9522 and Minghui 63. Quantitative real-time PCR showed that the 9 transcripts of OsJMJ718 present diverse dynamic expression patterns in different stages of rice reproductive development. The expression of OsJMJ718-TVX5 was higher than that of other tran- scripts. In summary, this work provides information for APA sequences of OsJMJ718, and the expression pattern analysis of these transcripts would also help further study of the mechanism of APA and biological function of OsJMJ718.

关 键 词：水稻 OsJMJ718 可选择性多聚腺苷酸化 3'RACE 表达模式 

分 类 号：Q78[生物学—分子生物学]