黑木耳YBS-3菌株ras启动子的克隆与分析

[目的]从黑木耳（Auricuraliaauricular）基因组中克隆内源Ⅻ启动子,为黑木耳利用基因工程培育优良品种提供启动子元件材料。[方法]利用PCR技术,以黑木耳菌株YBS一3基因组DNA为模版,克隆得到4条／ras启动子片段。采用启动子序列分析软件Place、Promoterprediction和TFSEARCHver．1．3对其进行序列结构分析。[结果]4条启动子片段均含有核心启动子序列,除典型的基本作用元件TATAbox和CAATbox外,还有许多其他重要作用元件如GATABOX、GCGCBOX和CCAATBOXl等,同时每条片段中均至少具有1个转录起始位点,且检测到HSF、AP-1、GCN4等多个转录因子结合位点。[结论]4条目的序列在理论上具有较强的启动子活性功能。

Cloning and Sequence Analysis of ras Promoter from Auricuralia auricular Strain YBS-3

[Objective] The ras promoters were cloned from genomic DNA of Auricuralia auricular so as to provide promoters for breeding better species by genetic technology.[Method] PCR was used to clone promoters with the genomic DNA of A.auricular strain YBS-3 as template,and then the sequences of four ras promoters were obtained.The analysis of promoter sequences was made by three promoter analysis software:Promoter prediction,Place and TFSEARCH ver.1.3.[Result] Four fragments contained core elements of promoter including TATA-box and CAAT-box,and many other important cis-elements such as GATA BOX,GCGC BOX,CCAAT BOX1,etc.At the same time,there was at least one transcriptional start site in these sequences.And several transcription factor binding sites were detected in these sequences.[Conclusion] Four promoter fragments were all having promoter function theoretically.