酵母钙转运缺陷株互补验证

服务介绍

K667酵母钙转运互补验证是一个经典实验体系，用来判断目标基因是否编码功能性钙（或其它阳离子）转运蛋白。其核心思路很简单：K667本身对高浓度钙敏感，生长会被抑制。如果转入了外源基因能“补上”缺失的钙转运功能，恢复它在高钙环境下的生长，就说明该基因很可能参与了钙的转运。

服务优势

▶可检测蛋白是否具有钙离子吸收转运活性，简单高效。

▶本司有钙离子转运活性检测所需的全部试剂、菌株和载体。

服务流程

文案展示

Arabidopsis calcium-dependent protein kinases 4/5/6/11 negatively regulate hydrotropism via phosphorylation of MIZU-KUSSEI1

期刊：The Plant Cell

CPK-mediated phosphorylation of MIZ1Ser14/36 is essential for facilitating ECA1 activity. A) Functional analysis of MIZ1 phosphorylated by CPKs on ECA1 activity in yeast. Growth of yeast strains on normal medium (lacking Ura and Leu, SD/−Ura/−Leu) and treatment medium (SD/−Ura/−Leu + 80 mM CaCl2) for 3 to 5 d. The strain used was K667 (Δpmc1vcx1cnb1), and K667 cells transformed with the empty pYES2 or P415 vectors  served as the negative control. From left to right, there is a 10-fold dilution series of yeast cultures. B and C) Growth curves of yeast cells were plotted against the A600 values in the SD/−Ura/−Leu (B) and SD/−Ura/−Leu + 80 mM CaCl2 nutrient solution (C). The A600 values of yeast cell growth were monitored from 12 to 72 h. The data are presented as mean±SD (n=3 biological replicates). One-way ANOVA and the LSD test for multiple comparisons were performed, and significant differences are indicated by letters (α=0.01), while “ns” indicates no significant differences.

适用领域

▶钙转运蛋白功能鉴定：鉴定植物、微生物、动物来源的候选钙转运蛋白基因的体内转运功能，明确目标基因是否具备介导钙离子跨膜吸收的核心活性，是基因功能验证的关键技术手段。

▶植物逆境生理研究：解析作物在缺钙、重金属胁迫等逆境下，钙转运蛋白在维持细胞内钙离子稳态中的作用；筛选耐低钙或钙高效利用的作物基因，为养分高效利用育种提供靶点。

▶微生物钙代谢机制解析：研究细菌、真菌等微生物的钙转运系统，明确钙转运蛋白在微生物适应钙匮乏环境、维持酶活性中的调控机制。

▶钙转运蛋白突变体/变体功能筛选：对钙转运蛋白的不同突变体、等位变体进行功能验证，评估氨基酸位点突变对转运活性的影响，助力蛋白结构 - 功能关系研究。