摘要
通过利用miRNA数据库和miRNA靶基因预测软件等生物信息学方法,筛选出靶基因与冷胁迫相关的miRNAs。在预测的柑橘冷胁迫相关miRNA中挑选5个miRNA进行表达量分析。提取不同冷处理时间的柑橘叶片总RNA,反转录成cDNA,用半定量RT-PCR的方法对5个miRNA的表达量分析。利用不同的RT-PCR技术,分别检测miRNA前体和其成熟体的拷贝数,发现在4°C冷处理不同时间后,多个miRNAs的表达量发生变化。研究和分析了柑橘中冷胁迫相关miRNAs的冷反应表达模式,为进一步明确这些miRNAs表达的调控机制以及柑橘中miRNAs在冷胁迫适应中的作用提供了依据。
The miRNAs of the citrus related to cold stress in target gene were selected by using bioinformatics method based on miRNA database and miRNA target gene prediction software.Five miRNAs were chosen from the predicted miRNAs related to cold stress to conduct expression analysis.Total RNA was isolated from citrus leaves which treated with different time of cold treatment,and was reverse transcribed to template cDNA.The expression of five miRNAs was analyzed by semi-quantitative RT-PCR.By using various RT-PCR methods,both miRNA precursor and mature miRNA were detected respectively,it found that different time after 4℃ cold treatment,the expression of many miRNAs had changed.The expression pattern of cold response of citrus miRNAs related to cold stress was studied and analyzed to provide references for further determining the regulatory mechanism of these citrus miRNAs and finding the effect of these citrus miRNAs in adaption to cold stress.
出处
《湖南农业科学》
2011年第5期13-16,共4页
Hunan Agricultural Sciences
基金
湖南省自然科学基金重点项目(08jj3066)
