拟南芥NaKR2超表达转基因株系的构建
Abstract: SODIUM POTASSIUM ROOT DEFECTIVE1 (NaKR1; previously called NPCC6) encodes a soluble metal binding protein that is specifically expressed in companion cells of the phloem. The nakr1-1 mutant phenotype includes high Na+, K+, Rb+, and starch accumulation in leaves, short roots, late flowering, and decreased long-distance transport of sucrose. Using traditional and DNA microarray-based deletion mapping, a 7-bp deletion was found in an exon of NaKR1 that introduced a premature stop codon. The mutant phenotypes were complemented by transformation with the native gene or NaKR1-GFP (green fluorescent protein) and NaKR1-β-glucuronidase fusions driven by the native promoter. NAKR1-GFP was mobile in the phloem; it moved from companion cells into sieve elements and into a previously undiscovered symplasmic domain in the root meristem. Grafting experiments revealed that the high Na+ accumulation was due mainly to loss of NaKR1 function in the leaves. This supports a role for the phloem in recirculating Na+ to the roots to limit Na+ accumulation in leaves. The onset of root phenotypes coincided with NaKR1 expression after germination. The nakr1-1 short root phenotype was due primarily to a decreased cell pision rate in the root meristem, indicating a role in root meristem maintenance for NaKR1 expression in the phloem.[1]
To research mechanisms of NaKR2 more deeply ,in this paper ,overexpression vector of NaKR1 gene was constructed .By the way of A grobacterium-mediated floral dipping methed , NaKR2-overexpression vector was transformed into Arabidopsis wild-type(Col) plants .the single cope homozygous were obtained by screening. These screened transgenic lines will be good materials for functional research of NaKR2.
毕业论文关键词：NAKR2；拟南芥； 超表达载体； PCR
Keyword：NAKR1； Arabidopsis；over-expression vector ；PCR
目 录
引言 4
1. 材料与方法 6
1.1. 实验材料 6
1.1.1. 植物材料 6
1.1.2. 菌株与载体 6
1.1.3. 试剂与仪器 6
1.2. 基本实验方法 7
1.2.1. 拟南芥种子处理 7
1.2.2. 大肠杆菌感受态细胞的制备 7
1.2.3. 质粒提取 7
1.2.4. 植物总 RNA 提取 8