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首页> 外文期刊>Infection and immunity >Molecular analysis of the serotyping antigens of Neisseria meningitidis.
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Molecular analysis of the serotyping antigens of Neisseria meningitidis.

机译：脑膜炎奈瑟氏球菌血清分型抗原的分子分析。

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Molecular approaches to the rapid analysis of the serotyping antigens of Neisseria meningitidis, the class 2 and 3 outer membrane proteins (OMPs), were developed, evaluated, and used to study 12 antigenic variants of these proteins. A primer set for the polymerase chain reaction (PCR) amplification of the genes encoding these antigens was devised. Low-stringency amplification of meningococcal chromosomal DNA with this primer set resulted in the amplification of two products from each strain, whereas at higher stringencies only one product was amplified in most strains. Southern hybridization techniques and restriction analyses were used to differentiate the PCR products amplified at high stringencies from strains expressing class 2 or class 3 OMPs; these PCR products were further characterized by the determination of their nucleotide sequences, confirming that they represented the amplified class 2 and class 3 OMP genes. Analyses of these and other nucleotide sequences enabled the construction of a phenogram illustrating the interrelationships between Neisseria OMP genes. The comparative analysis of deduced amino acid sequences revealed conserved and variable regions of the proteins; the latter probably correspond to surface loops on the protein and hence are potentially exposed to the immune system. Further analyses of the primary structures of these related porins from Neisseria species enabled construction of models of the secondary structure of these antigens and comparison of these models with those previously published. The methods reported in the present work are rapid reproducible procedures for the analysis of antigenic variants of these proteins.
机译：已开发，评估和评估了脑膜炎奈瑟氏球菌血清型抗原的2类和3类外膜蛋白（OMP）的分子生物学方法，并用于研究这些蛋白的12种抗原变体。设计用于编码这些抗原的基因的聚合酶链反应（PCR）扩增的引物组。用该引物组对脑膜炎球菌染色体DNA进行低严格扩增会导致每个菌株扩增出两种产物，而在更高严格性下，大多数菌株只会扩增一种产物。使用Southern杂交技术和限制性酶切分析，将高严格扩增的PCR产物与表达2类或3类OMP的菌株区分开来。通过确定其核苷酸序列，进一步表征了这些PCR产物，证实它们代表了扩增的2类和3类OMP基因。对这些核苷酸序列和其他核苷酸序列的分析使得能够构建表示奈瑟氏菌OMP基因之间相互关系的物候图。推导的氨基酸序列的比较分析揭示了蛋白质的保守和可变区。后者可能对应于蛋白质上的表面环，因此可能暴露于免疫系统。对来自奈瑟氏球菌属的这些相关孔蛋白的一级结构的进一步分析使得能够构建这些抗原的二级结构的模型并将这些模型与先前公开的那些进行比较。在本工作中报道的方法是用于分析这些蛋白质的抗原变体的快速可重复的程序。

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