参环毛蚓MT-2基因及启动子的克隆研究

发布时间：2018-03-05 18:07

本文选题：参环毛蚓　切入点：重金属富集　出处：《中草药》2016年16期 　论文类型：期刊论文

【摘要】：目的揭示参环毛蚓MT-2基因的转录调控机制,为今后采用基因工程技术改良参环毛蚓重金属富集性状奠定基础。方法根据已知的MT-2c DNA序列,设计特异引物,通过PCR扩增参环毛蚓MT-2基因的编码区基因组序列,扩增产物经测序后自行比对分析。基于该序列设计3条特异引物,采用基因步移技术克隆其上游的启动子序列,同时运用Promoter Prediction在线软件分析预测其顺式元件。结果经PCR扩增、测序后获得2 826 bp MT-2基因编码区序列,将该序列与已知MT-2 c DNA序列(登录号为KC787373.1)进行比对,发现MT-2编码区由4个外显子和4个内含子组成。基因步移技术克隆得到1 534 bp的启动子序列,经Promoter Prediction在线软件分析,结果显示该序列不仅含有TATA-box、CAAT-box等核心启动子软件,还具有3个特异响应重金属参与调控MT-2表达的MRE元件。结论参环毛蚓的MT-2基因能被重金属诱导表达,MT-2基因转录水平的金属调控是通过MT-2基因启动子区的金属调控元件MRE来实现的。
[Abstract]:Objective to elucidate the transcriptional regulation mechanism of MT-2 gene and lay a foundation for the improvement of heavy metal enrichment by genetic engineering. Methods based on the known MT-2c DNA sequence, specific primers were designed. The coding region of MT-2 gene was amplified by PCR, and the amplified product was sequenced and analyzed. Based on the sequence, three specific primers were designed, and the upstream promoter sequence was cloned by gene step technique. At the same time, the cis element was predicted by Promoter Prediction online software. Results after PCR amplification, the sequence of 2826 BP MT-2 gene coding region was obtained, and the sequence was compared with the known MT-2 c DNA sequence (accession number KC787373.1). It was found that the coding region of MT-2 was composed of 4 exons and 4 introns. The promoter sequence of 1 534 BP was cloned by gene step technique and analyzed by Promoter Prediction online software. The results showed that the sequence contained not only the core promoter software such as TATA-box CAAT-box, but also the core promoter software such as TATA-box. There are also three MRE elements that specifically respond to heavy metals to regulate the expression of MT-2. Conclusion the metal regulation of the transcription level of the MT-2 gene can be induced by heavy metal in the promoter region of the MT-2 gene through the metal regulation of the promoter region of the MT-2 gene. Component MRE to achieve.
【作者单位】：广州中医药大学中药学院;湖北中医药大学药学院;
【基金】：国家自然科学基金项目(30772741) 广东省科技计划项目(2009B060700006) 广州市科技计划项目(2010Y1-C981)
【分类号】：S865.4;Q78

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