拟南芥耐镉基因的鉴定和玉米韧皮部蛋白的提取
时间:2018-08-26 19:47 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：镉是一种重金属污染物,对植物具有显著的毒害作用。植物中积累的镉可通过食物链进入人体，给人类健康带来潜在危害。本实验利用镉敏感酵母突变株Δycf1，通过酵母异源功能互补实验和酵母生长曲线测定，验证了拟南芥VPS29基因的耐镉性。玉米对多种环境因子的胁迫均表现出较强的抗性，是广泛用于重金属污染研究的重要农作物。韧皮部蛋白质在植物大分子运输系统中起很重要的标记作用。在镉处理的玉米中，采用切割渗出的方法提取韧皮部蛋白，冷冻干燥成粉，利用SDS-PAGE电泳技术来检测提取的蛋白。27421 毕业论文关键词：镉；酵母异源功能互补；VPS29；韧皮部蛋白；玉米 Analysis of the tolerance of Arabidopsis vacuolar sorting protein to cadmium and extracting of phloem proteins of Maize Abstract：Cadmiun(Cd) is one of the toxic heavy metals,which has obvious toxic effects on plants and represents as potential threat to human health through food chain.Maize is one of the crop which is extensively planted in the north area of our country and it can be resistent to many wicked environmental factors.In this study,we used cadmium sensitive yeast mutant Δycf1,according to the results of yeast complementary experiment and the transformed yeast growth curve,we identitied that Δycf1 strains transformed with Arabidopsis VPS29 gene showed tolerance to cadmium stress.Maize is one of the C4 plant, has rapid growth and enormous output.Maize can be resistent to many wicked environmental factors, has great potential in the restoration of plant and is widely used in the study of heavy metal pollution.Maize under heavy Cd stress inbihits plant growth,causes alterations in morphological and physiological disorders,affects several plant metabolic processes.Phloem protein palys important mark rloe in the transportation of macromolecular.The cutting seeping method is reported useful in collecting sap of phloem of the maize which is under heavy Cd stress.There are several high throughput proteomics approaches used to study proteomics ,sodium dodecyl sulphate polyacrylamide gel electrophoresis(SDS-PAGE) is usually used to detect the protein purity of the liquid extracted from maize phloem using the cutting seeping method .After the freeze drying we used SDS-PAGE to analyze the phloem protein. Key words: Cadmiun;yeast complementary experiment;VPS29; phloem protein;maize 目 录 摘要 1 关键词 1 Abstract 1 Key words 1 引言（或绪论） 1 1 材料与方法 3 1.1 实验材料 3 1.1.1 酵母菌株与载体 3 1.1.2 植物材料 3 1.2 仪器与试剂 3 1.3 培养基 3 1.4 实验方法 3 1.4.1酵母感受态制备与转化 3 1.4.2 确定镉的浓度 4 1.4.3 菌落PCR 4 1.4.4 PCR扩增目的基因 4 1.4.5 目的基因的回收纯化 5 1.4.6目的基因连接T载体 5 1.4.7 酵母表达载体的构建 5 1.4.8 酵母感受态的制备 5 1.4.9 酵母感受态细胞转化 5 1.4.10 酵母异源功能互补实验 5 1.4.11酵母生长曲线测定实验 6 1.4.12玉米幼苗的培养及韧皮部蛋白的提取 6 1.4.13 SDS-PAGE分析 6 2 结果与分析 7 2.1 拟南芥VPS29基因耐镉性的验证 7 2.1.1 酵母表达载体的构建 7 2.1.2 酵母异源功能互补实验验证鉴定拟南芥VPS29基因耐镉性 7 (责任编辑：qin) |