烟草NtGSH2基因的克隆及表达特性分析

发布时间：2018-05-07 20:03

本文选题：基因表达 + 谷胱甘肽　；参考：《基因组学与应用生物学》2017年05期

【摘要】：谷胱甘肽(glutathione,GSH)是生物体内的非蛋白巯基类化合物,具有抗氧化、抗衰老和重金属解毒等重要的生理功能。本研究从烟草(Nicotiana tabacum L.)中克隆到谷胱甘肽合成酶基因NtGSH2,该基因编码555个氨基酸残基的蛋白质。氨基酸序列比对和进化树分析发现,NtGSH2与其他植物同源蛋白具有较高的一致性,且与枸杞(Lycium chinense)LcGSHS/LcGS和番茄(Solanum lycopersicum)SlGSH2归为一类,亲缘关系最近。组织表达模式分析表明,NtGSH2基因在烟草的根、茎、叶、花和种子中均表达,其中在花中的表达量最高,在种子中的表达量最低。实时荧光定量PCR检测表明,NtGSH2受CdCl_2和ZnCl_2诱导表达。将NtGSH2连接到原核表达载体pET32a上,转化获得含有NtGSH2基因的大肠杆菌BL21(DE3)工程菌并成功表达目的蛋白。该研究结果为进一步分析NtGSH2基因功能及谷胱甘肽的生物合成提供了一定的理论依据。
[Abstract]:Glutathione glutathione (GSH) is a nonprotein sulfhydryl compound which has important physiological functions such as antioxidation, anti-aging and detoxification of heavy metals. In this study, Nicotiana tabacum L. The glutathione synthase gene NtGSH _ 2, which encodes 555 amino acid residues, was cloned. Amino acid sequence alignment and phylogenetic tree analysis showed that NtGSH _ 2 had high consistency with other plant homologous proteins, and was closely related to Lycium chinense)LcGSHS/LcGS and Solanum lycopersicum)SlGSH2 of Lycium barbarum. Tissue expression pattern analysis showed that NtGSH2 gene was expressed in roots, stems, leaves, flowers and seeds of tobacco, among which the highest expression was in flowers and the lowest in seeds. Real time fluorescence quantitative PCR assay showed that NtGSH2 was induced by CdCl_2 and ZnCl_2. NtGSH2 was ligated into the prokaryotic expression vector pET32a and transformed into E. coli BL21 (DE3) containing NtGSH2 gene and the target protein was successfully expressed. The results provide a theoretical basis for further analysis of NtGSH2 gene function and glutathione biosynthesis.
【作者单位】：西南科技大学生命科学与工程学院;西南科技大学核废物与环境安全国防重点实验室;
【基金】：国家核设施退役及放射性废物治理科研重点项目(No.14ZG6101) 国家自然科学基金(No.31500205) 西南科技大学核废物与环境安全国防重点学科实验室预研基金(No.15yyhk09);西南科技大学重点科研平台专职科研创新团队基金(No.14tdhk02)共同资助
【分类号】：Q943.2

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