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郭艳, 张会, 简曙光, 夏快飞, 张美. 厚藤cDNA文库的构建和重金属镉耐受相关基因的筛选[J]. 植物科学学报, 2017, 35(3): 372-378. DOI: 10.11913/PSJ.2095-0837.2017.30372
引用本文: 郭艳, 张会, 简曙光, 夏快飞, 张美. 厚藤cDNA文库的构建和重金属镉耐受相关基因的筛选[J]. 植物科学学报, 2017, 35(3): 372-378. DOI: 10.11913/PSJ.2095-0837.2017.30372
Guo Yan, Zhang Hui, Jian Shu-Guang, Xia Kuai-Fei, Zhang Mei. Construction of a cDNA library of Ipomoea pes-caprae L.and screening for cadmium tolerant genes[J]. Plant Science Journal, 2017, 35(3): 372-378. DOI: 10.11913/PSJ.2095-0837.2017.30372
Citation: Guo Yan, Zhang Hui, Jian Shu-Guang, Xia Kuai-Fei, Zhang Mei. Construction of a cDNA library of Ipomoea pes-caprae L.and screening for cadmium tolerant genes[J]. Plant Science Journal, 2017, 35(3): 372-378. DOI: 10.11913/PSJ.2095-0837.2017.30372

厚藤cDNA文库的构建和重金属镉耐受相关基因的筛选

Construction of a cDNA library of Ipomoea pes-caprae L.and screening for cadmium tolerant genes

  • 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ycf1△,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ycf1△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IpPCSIpMT,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,as a coastal beach halophyte with great ecological,ornamental,and medicinal value,has the capacity to enrich heavy metal cadmium and can be used in phytoremediation.We constructed a cDNA library for expression in yeast with tissues of Ipomoea pes-caprae by gateway technology,and then introduced the library plasmids into cadmium-sensitive yeast mutant strain,ycf1△,and screened the library through the FOX gene hunting system based on functional screening in yeast.We obtained two full-length cDNAs of Ipomoea pes-caprae,which could enhance the Cd tolerance of ycf1△.Sequence analyses indicated that these two cDNAs encoded phytochelatin synthase (IpPCS) and metallothionein (IpMT).This research lays a good foundation for further clarifying the molecular mechanism of plants responding to cadmium,and provides a reference cDNA library for functional screening to clone other stress-related genes in Ipomoea pes-caprae.

     

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