脑膜炎奈瑟菌lpdA基因在大肠杆菌中的表达、P64k蛋白的蛋白质序列分析及免疫学特性研究

Expression in Escherichia coli of the lpdA gene, protein sequence analysis and immunological characterization of the P64k protein from Neisseria meningitidis.

作者信息

Guillén G, Alvarez A, Silva R, Morera V, González S, Musacchio A, Besada V, Coizeau E, Caballero E, Nazabal C, Carmenate T, González L J, Estrada R, Támbara Y, Padrón G, Herrera L

机构信息

Division of Vaccines, Centro de Ingeniería Genética y Biotecnología, La Habana, Cuba.

出版信息

Biotechnol Appl Biochem. 1998 Jun;27(3):189-96.

PMID:9664678

Abstract

By making use of recombinant DNA technology it is possible to characterize meningococcal outer membrane proteins (OMPs) capable of stimulating a host immune response. The lpdA gene, which codes for an OMP (P64k) from Neisseria meningitidis, was cloned in Escherichia coli. The recombinant protein was recognized by sera from patients convalescing from meningococcal disease. The monoclonal antibodies obtained against the recombinant protein recognized the natural protein on a Western blot, and monoclonal antibody 114 was assayed in ELISA with a panel of 85 N. meningitidis strains. The protein was recognized in 81 strains (95.3%); the strains that were not recognized were neither epidemic nor isolated from systemic disease. The complete amino acid sequence of P64k was obtained by automatic sequencing and MS.

摘要

利用重组DNA技术能够鉴定出可刺激宿主免疫反应的脑膜炎球菌外膜蛋白（OMPs）。编码脑膜炎奈瑟菌一种OMP（P64k）的lpdA基因被克隆到大肠杆菌中。重组蛋白能被脑膜炎球菌病康复期患者的血清识别。针对重组蛋白获得的单克隆抗体在蛋白质印迹法中能识别天然蛋白，并且用一组85株脑膜炎奈瑟菌菌株在酶联免疫吸附测定（ELISA）中检测了单克隆抗体114。在81株菌株（95.3%）中识别出了该蛋白；未被识别的菌株既非流行菌株，也不是从全身性疾病中分离得到的。通过自动测序和质谱分析法获得了P64k的完整氨基酸序列。