Insertion mutagenesis of the Pseudomonas aeruginosa phosphate-specific porin OprP
Insertion mutagenesis of the Pseudomonas aeruginosa phosphate-specific porin OprP
复制标题
铜绿假单胞菌磷酸盐特异性孔蛋白 OprP 的插入诱变
DOI：
10.1128/jb.177.17.4914-4920.1995
复制
发表时间：
1995
影响因子：
3.2
通讯作者：
Robert E. W. Hancock
中科院分区：
文献类型：
--
作者：
A. Sukhan;Robert E. W. Hancock
对编码铜绿假单胞菌磷酸盐特异性孔蛋白OprP的基因进行接头诱变和表位插入诱变。 13 个接头突变体基因中的 9 个表达的蛋白质水平与野生型基因获得的水平相当。使用 OprP 特异性抗血清进行间接免疫荧光显示，这些突变蛋白正确暴露在细胞表面。四个接头突变体基因表达的蛋白质水平降低，在细胞表面无法检测到。来自疟疾寄生虫恶性疟原虫的环子孢子形式的外源表位被克隆到 13 个突变基因中的 12 个的连接位点中。所得表位插入突变基因中的七个表达表面暴露蛋白。其中两个突变基因在表面可及区域呈现外源表位，使用疟疾表位特异性单克隆抗体通过间接免疫荧光进行评估。这些实验的数据用于创建 OprP 单体的拓扑模型。
The gene encoding the Pseudomonas aeruginosa phosphate-specific porin OprP was subjected to both linker and epitope insertion mutageneses. Nine of the 13 linker mutant genes expressed protein at levels comparable to those obtained with the wild-type gene. These mutant proteins were shown, by indirect immunofluorescence with an OprP-specific antiserum, to be properly exposed at the cell surface. Four of the linker mutant genes expressed protein at reduced levels which were not detectable at the cell surface. A foreign epitope from the circumsporozoite form of the malarial parasite Plasmodium falciparum was cloned into the linker sites of 12 of the 13 mutant genes. Seven of the resultant epitope insertion mutant genes expressed surface-exposed protein. Two of these mutant genes presented the foreign epitope at surface-accessible regions as assessed by indirect immunofluorescence with a malarial epitope-specific monoclonal antibody. The data from these experiments were used to create a topological model of the OprP monomer.
铜绿假单胞菌外膜的蛋白质 D2 通道具有碱性氨基酸和肽的结合位点。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Trias,J;Nikaido,H
通讯作者：
Nikaido,H