摘要
采用特异反转录引物,构建莱茵衣藻microRNA(miRNA)的cDNA文库.选用U4核仁小分子RNA(small nucleolar RNA,snoRNA)作为内参,用SYBR green RT-PCR对3种与莱茵衣藻缺硫胁迫反应相关的miRNA进行检测,利用在线平台软件Web MicroRNAs Designer(WMD3)对miRNA靶基因进行预测.结果表明,在缺硫胁迫下,3种miRNA(miRNA1145.2、miRNA1146和miRNA1158)的表达水平均明显上调,与正常培养的莱茵衣藻表达miRNA的相对丰度比值分别为3.11、2.38和3.67.靶基因预测分析表明,miRNA1145.2能影响脂肪酸氧化反应,miRNA1146与辅酶Q代谢相关,miRNA1158可能参与磷酸戊糖途径调控.
Two small RNA libraries,which responded to sulfur-replete and sulfur-deprivation condition,were generated by using stem-loop primers in Chlamydomonas reinhardtii.Three microRNAs（miRNA1145.2,miRNA1146 and miRNA1158） were quantified by using SYBR green reverse transcription polymerase chain reaction（RT-PCR） detection with U4 snoRNA as internal reference gene.Their target genes were predicted by the Web MicroRNAs Designer（WMD3）.The results showed that all three microRNAs expressions were up-regulated under sulfur deprivation in Chlamydomonas reinhardtii.The ratio of relative abundance with/without sulfur deprivation for miRNA1145.2,miRNA1146 and miRNA1158 were 3.11,2.38 and 3.67,respectively.The miRNA1145.2 was able to target the thiolase gene to influence the fatty acid metabolism.The miRNA1146 targeted the genes for ubiquinone/menaquinone biosynthesis methyltransferase to control the ubiquinone metabolism.The miRNA1158 targeted the 6-phosphogluconate dehydrogenase genes to regulate the pentose phosphate pathway.Results show that the photosynthetic metabolism could be changed by the expression of endogenous miRNAs to regulate their target genes.And we can also design miRNAs to regulate any specific genes that catch our attention.
出处
《深圳大学学报（理工版）》
EI
CAS
北大核心
2011年第3期237-242,共6页
Journal of Shenzhen University(Science and Engineering)
基金
国家自然科学基金资助项目(31070323)~~
