海滨雀稗PvHMA3基因克隆及其序列分析
毕业论文关键词：镉污染，海滨雀稗，PvHMA3，蛋白修饰
CLONING AND SEQUENCE ANALYSIS OF PvHMA3 GENE FROM SEASHORE PASPALUM
As one of the most toxic heavy metal elements,cadmium is an important stress factor affecting plant growth and ecological safety.The use of cadmium - tolerant turfgrass for soil heavy metal remediation has the advantages of green,economic and environmental protection,inheritance of cadmium - resistant turfgrass resources and analysis of cadmium-resistant molecular mechanism can speed up the process of cadmium-resistant breeding of turfgrass.The previous research has found Paspalum vaginatum is the strongest cadmium-resistant warm season turfgrass,but its cadmium-resistant molecular mechanism is unclear.It has been found that heavy metal ATPase3 (HMA3) regulates plant cadmium tolerance;in this study, PvHMA3 gene was cloned from paspalum viridis, and the sequence analysis, transmembrane analysis and protein modification site were detected by bioinformatics software.The implementation of this study will provide excellent gene resources for cadmium resistance breeding of turfgrass, and lay the foundation for further analysis of the post-translational modification mechanism of HMA protein.
Key words:soil cadmium pollution,seashore paspalum,PvHMA3,protein modification
目 录
摘要 1
关键词 1
ABSTRACT 1
KEY WORDS 1
1材料与方法 2
1.1材料 2
1.1.1植物材料 2
1.1.2载体 3
1.1.3试剂及试剂盒 3
1.1.4培养基 3
1.2方法 3
1.2.1序列同源搜索及引物的设计 3
1.2.2海滨雀稗总RNA的提取 3
1.2.3海滨雀稗RNA反转录cDNA 3
1.2.4海滨雀稗PvHMA3基因的克隆 4
1.2.5构建植物表达载体 5
1.2.6菌落PCR 6
1.2.7质粒小提试剂盒方法 6
2结果与分析 7
2.1PvHMA3基因cDNA全长的克隆 7
2.2PvHMA3的序列分析 7
2.3PvHMA3 蛋白跨膜结构域 10
2.4PvHMA3蛋白修饰位点的预测 11
3讨论 11
致谢 12
参考文献 13
海滨雀稗PvHMA3基因克隆及其序列分析
重金属污染已经成为现阶段严重威胁的环境的全球性问题之一。在所有的重金属污染物中，镉（Cd）是最具有植物毒性的物质之一[1]，其一方面通过食物链的富集作用进入人体，严重威胁人类健康[2]，同时，土壤中过量的镉会抑制植物生长、破坏细胞结构、降低光合作用以及酶活性，从而影响植物产量和质量。由此可见，土壤中重金属镉的生态修复显得尤为重要。据目前研究来看，土壤镉污染的修复技术具体包括了两大类，即物理化学修复和生物修复。植物修复是指利用一些特定植物的某种特殊生理机制，以达到除去污染土壤和废水中重金属的目的，并以其绿色净化、对环境不造成二次污染、投资成本低、应用方便等诸多优势成为研究生物修复的热点。植物修复的前提是培育耐镉性强或镉富集的材料，而阐明植物应对镉胁迫的生理及分子机制是培育耐镉新种质的基础。然而，目前的一些耐镉机制更多集中在拟南芥、水稻、小麦等耐镉性差的植物中，耐镉性强或镉富集植物的耐镉机制尚少，有待进一步解析。