Mapping of Chlamydia trachomatis proteins by immobiline-polyacrylamide two-dimensional electrophoresis: Spot identification by N-terminal sequencing and immunoblotting
Mapping of Chlamydia trachomatis proteins by immobiline-polyacrylamide two-dimensional electrophoresis: Spot identification by N-terminal sequencing and immunoblotting
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DOI：
10.1002/elps.1150170130
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发表时间：
1996-01-01
期刊：
影响因子：
2.9
通讯作者：
Pallini, V
中科院分区：
文献类型：
--
作者：
Bini, L;SanchezCampillo, M;Pallini, V
通过二维凝胶电泳在第一维非线性宽范围固定化 pH 梯度和第二维聚丙烯酰胺梯度凝胶上分离来自纯化的沙眼衣原体原体的蛋白质。使用该系统获得的地图具有高度可重复性，并且分辨率约为。 600 个名额。通过使用特异性抗体和/或 N 端氨基酸测序的免疫印迹分析，我们建立了许多所述衣原体蛋白的图谱位置，例如主要外膜蛋白 (MOMP)、60 kDa 富含半胱氨酸的外膜蛋白 (OMP2)、DnaK 样、GroEL 样和巨噬细胞感染增强剂 (MIP) 样蛋白、质粒编码的 pgp3 蛋白、两种核糖体蛋白（S1 和 L7/L12），以及蛋白质延伸因子 EF-Tu。其他无法进行基因分配的蛋白质已通过三个参数（M(r)、pI 和 N 末端序列）进行鉴定。这项工作为未来逐步编译衣原体蛋白基因组连锁数据库提供了初步基础。
Proteins from purified elementary-bodies of Chlamydia trachomatis were separated by two-dimensional gel electrophoresis on nonlinear wide-range immobilized pH gradients in the first dimension and polyacrylamide gradient gels in the second dimension. The maps obtained with this system are highly reproducible and resolve ca. 600 spots. By using immunoblot analysis with specific antibodies and/or N-terminal amino acid sequencing, we established the map positions of a number of described chlamydial proteins, such as the major outer membrane protein (MOMP) the 60 kDa cystein-rich outer membrane protein (OMP2), the DnaK-like, GroEL-like, and macrophage infectivity potentiator (MIP)-like proteins, the plasmid-encoded pgp3 protein, two ribosomal proteins (S1 and L7/L12), and the protein-elongation factor EF-Tu. Other proteins, for which gene assignment was not possible, have been identified by three parameters (M(r), pI and N-terminal sequence). This work provides a preliminary basis for a future and progressive compilation of a genome-linked database of chlamydial proteins.