Quantitative determination of FtsA at different growth rates in Escherichia coli using monoclonal antibodies
Quantitative determination of FtsA at different growth rates in Escherichia coli using monoclonal antibodies
复制标题
使用单克隆抗体定量测定大肠杆菌不同生长速率下的 FtsA
作者：
Haichao Wang;R. Gayda
FtsA是大肠杆菌中一种必需的细胞分裂蛋白。它的合成量低，使其功能的研究变得困难。过表达FtsA基因后，通过溶解细胞包涵体获得部分纯化的FtsA蛋白，以提高单克隆抗体。用该FtsA蛋白片段免疫小鼠，将其脾脏细胞与Sp2/0‐AG14小鼠骨髓瘤细胞融合。对杂交细胞进行筛选，通过酶联免疫吸附试验和Western blotting鉴定两个克隆为FtsA单克隆抗体的产生者。使用这些单克隆抗体的定量分析表明，每个细胞的平均FtsA分子数在50到200之间。然而，在很宽的生长速率范围内，FtsA蛋白与细胞总蛋白的标准化浓度是恒定的。这一发现与FtsA蛋白作为鼻中隔化学计量成分的假设作用一致。
FtsA is an essential cell division protein in Escherichia coli. Its synthesis in low amounts makes the investigation of its functions difficult. Partially purified FtsA protein was obtained by solubilizing cellular inclusion bodies after overexpression of the ftsA gene for the purpose of raising monoclonal antibodies. Mice were immunized with this FtsA protein fraction and their spleen cells were fused to Sp2/0‐AG14 mouse myeloma cells. Hybrid cells were screened and two clones were positively identified as FtsA monoclonal antibody producers by enzyme‐linked immunosorbent assay and Western blotting. A quantitative assay using these monoclonal antibodies indicated that the average number of FtsA molecules per cell to be between 50 and 200. However, the concentration of FtsA protein normalized to total cell protein was constant over a wide range of growth rates. This finding is in agreement with the hypothesized role of FtsA protein as a stoichiometric component of the septum.
大肠杆菌细胞分裂必需基因 ftsZ 的核苷酸序列。
影响因子：
3.5
作者：
Yi,QM;Lutkenhaus,J
通讯作者：
Lutkenhaus,J
细胞分裂基因 ftsQ、ftsA 和 ftsZ 的结构和表达。
影响因子：
5.6
作者：
Yi,QM;Rockenbach,S;WardJr,JE;Lutkenhaus,J
通讯作者：
Lutkenhaus,J