大白菜KIN基因的表达及其pre-mRNA加工机制研究

项目名称： 大白菜KIN基因的表达及其pre-mRNA加工机制研究

项目编号： No.31301788

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 陶鹏

作者单位： 浙江省农业科学院

项目金额： 23万元

中文摘要： 持续的冻害会引起大白菜产量和品质的下降，抗冻基因的诱导表达对保护其渡过低温环境起着关键作用，转录后加工是调控抗冻基因表达的重要阶段。本项目以白菜全基因组测序为基础，以抗/不抗冻的大白菜为材料，研究抗冻基因KIN的表达及其pre-mRNA加工特点。通过RACE结合PAGE技术，分析KIN基因的3'UTR的多态性，鉴定多聚腺苷酸化位点及频率。通过特异引物进行PCR，鉴定KIN的不同剪接位点。比较分析抗冻/不抗冻大白菜种质间KIN基因的pre-mRNA加工的差异，找出与大白菜抗冻性密切相关的主要pre-mRNA加工特征。在此基础上，对大白菜在不同激素和胁迫处理下KIN基因的pre-mRNA加工特点深入分析，确定KIN基因发挥抗冻功能的主要转录后加工产物。该研究结果对了解大白菜抗逆基因转录后加工机制与胁迫之间的关系具有重要意义，为下步开展KIN的pre-mRNA加工产物的下游生物学功能研究提供基础

中文关键词： 大白菜；KIN；选择性剪接；选择性多聚腺苷酸化；机制

英文摘要： Low temperature is one of the most important abiotic factors limiting productivity and quality of Brassica campestris L.ssp.pekinensis. Cold-regulated (COR) genes can be induced to express by cold stress, and play a key role in conferring freezing/cold tolerance of plants. Post-transcriptional regulation is one of the important stages in regulating the expression of COR genes. Due to whole genome sequencing of Brassica rapa (Chiifu-401), it is convenient and important for analyzing the expression and pre-mRNA processing mechanism of the cold-regulated KIN genes from the antifreeze / not antifreeze materials of Brassica campestris L.ssp.pekinensis. By RACE combining with PAGE, we can analyzed the length polymorphism of 3'UTR of the KIN transcripts for checking the sites and frequency of polyadenylation of the KIN genes from Brassica campestris L.ssp.pekinensis. Further, the different spliced transcripts of the KIN genes, produced from the same pre-mRNA by alternative splicing, are obtained and sequenced for analyzing the spliced sites of the KIN genes by PCR with specific primer sets. We analyzed comparatively the differences of polyadenylation of the KIN genes in pre-mRNA processing between different varieties of Brassica campestris L.ssp.pekinensis, and characterized the pre-mRNA processing obviously related to

英文关键词： Chinese cabbage；KIN；Alternative splicing；Alternative polyadenylation；mechanism