Sperker B, Hu P, Herrmann R
Department of Microbiology, University of Heidelberg, Germany.
Mol Microbiol. 1991 Feb;5(2):299-306. doi: 10.1111/j.1365-2958.1991.tb02110.x.
Gene P1 of Mycoplasma pneumoniae, which codes for a major adhesin, is flanked by two sequences with open reading frames designated ORF4 and ORF6 (Inamine et al., 1988b). In order to identify proteins translated from those ORFs, gene fusions between the N-terminus of the RNA replicase of the Escherichia coli bacteriophage MS2 and selected regions of ORF4 and ORF6 were constructed. The corresponding fusion proteins synthesized in Escherichia coli were used to immunize mice. Antisera directed against ORF4-related sequences did not recognize M. pneumoniae antigens in Western blot analysis, but antisera directed against ORF-6-derived fusion proteins reacted with two M. pneumoniae proteins of 40 kDa and 90 kDa. In addition, some of the antisera also recognized proteins that formed in a sodium dodecyl sulphate/polyacrylamide gel a protein ladder between 115 and 145 kDa.
肺炎支原体的基因P1编码一种主要黏附素，其两侧有两个带有开放阅读框的序列，分别命名为ORF4和ORF6（稻岭等人，1988b）。为了鉴定从这些开放阅读框翻译出的蛋白质，构建了大肠杆菌噬菌体MS2的RNA复制酶N端与ORF4和ORF6选定区域之间的基因融合体。在大肠杆菌中合成的相应融合蛋白用于免疫小鼠。在蛋白质印迹分析中，针对ORF4相关序列的抗血清未识别出肺炎支原体抗原，但针对ORF-6衍生融合蛋白的抗血清与两种分子量分别为40 kDa和90 kDa的肺炎支原体蛋白发生反应。此外，一些抗血清还识别在十二烷基硫酸钠/聚丙烯酰胺凝胶中形成的分子量在115至145 kDa之间的蛋白梯带中的蛋白质。
