厚藤cDNA文库的构建和重金属镉耐受相关基因的筛选

发布时间：2018-11-07 07:27

【摘要】：厚藤(Ipomoea pes-caprae(L.)Sweet)是一种具有重要生态、观赏及药用价值的沙滩植物,对重金属镉(Cd)具有一定的富集能力,可作为Cd污染滨海地区的修复植物进行引种栽植和利用。本研究通过Gateway技术构建厚藤的cDNA文库,将该文库质粒转化酵母对Cd敏感的突变株ycf1Δ,采用全长cDNA过表达基因捕获系统(FOX)筛选厚藤重金属Cd胁迫耐受相关基因,并采用酵母互补实验进行基因的功能验证。本研究获得了2个能够恢复ycf1Δ对Cd敏感表型的重组质粒,经测序分析,该重组质粒包含的cDNA全长序列分别对应厚藤植物螯合肽合成酶基因(phytochelatin synthase)和金属硫蛋白基因(metallothionein),分别将其命名为Ip PCS和Ip MT,通过功能分析,初步认定该基因为编码Cd耐受和解毒相关蛋白的候选基因。
[Abstract]:Ipomoea pes-caprae (L.) Sweet) is a kind of beach plant with important ecological, ornamental and medicinal value. It has certain enrichment ability to cadmium (Cd). It can be used as a repair plant in coastal areas polluted by Cd. In this study, the cDNA library was constructed by Gateway technique. The plasmid was transformed into the mutant ycf1 螖, which was sensitive to Cd. The full-length cDNA overexpression gene capture system (FOX) was used to screen the genes related to Cd stress tolerance. The function of the gene was verified by yeast complementary experiment. In this study, two recombinant plasmids capable of restoring ycf1 螖 sensitive phenotype to Cd were obtained. By sequencing, the full length cDNA sequence of the recombinant plasmid was found to correspond to the chelate peptide synthase gene (phytochelatin synthase) and the metallothionein gene (metallothionein), respectively. They were named Ip PCS and Ip MT, respectively by functional analysis and identified as candidate genes encoding Cd tolerance and detoxification related proteins.
【作者单位】：中国科学院华南植物园广东省应用植物学重点实验室;中国科学院大学;
【基金】：国家自然科学基金项目(31271702,31570257) 中国科学院A类战略性先导科技专项(XDA13020500) 国家重点研发计划项目(2016YFC1403002) “十二五”农村领域国家科技计划项目(2015BAL04B04)资助~~
【分类号】：Q943.2

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