Watanabe K, Onoe T, Ozeki M, Shimizu Y, Sakayori T, Nakamura H, Yoshimura F
Department of Health Education, Aichi University of Education, Japan.
Microbiol Immunol. 1996;40(10):725-34. doi: 10.1111/j.1348-0421.1996.tb01133.x.
The downstream DNA region of the fimbrilin gene (fimA), which encodes the major subunit protein of Porphyromonas gingivalis fimbriae, was fully sequenced. Gene products, expressed from this region in Escherichia coli, were purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their partial amino acid sequences were determined to verify open reading frames (ORFs) found in the region by DNA sequencing. Four ORFs, designated ORF1, ORF2, ORF3 and ORF4, were found in the 5.8-kb PstI fragment downstream from fimA, which was previously cloned and partially characterized by Yoshimura, Takahashi, Hibi, Takasawa, Kato, and Dickinson (Infect. Immun. 61: 5181-5189, 1993). The direction of transcription of all the ORFs was the same as that of fimA. The 50 and 80 kDa encoded proteins, ORF2 and ORF3, respectively, have been reported to be minor components associated with fimbriae. The 15 and 19 kDa proteins, ORF1 and ORF4, respectively, have been expressed in E. coli but not identified in P. gingivalis. However, all the gene products of the ORFs, expressed in E. coli, appeared to contain intact signal peptides based on their N-terminal amino acid sequences.
对编码牙龈卟啉单胞菌菌毛主要亚基蛋白的菌毛蛋白基因(fimA)的下游DNA区域进行了全序列测定。在大肠杆菌中从该区域表达的基因产物通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行纯化,并测定其部分氨基酸序列,以验证通过DNA测序在该区域发现的开放阅读框(ORF)。在fimA下游的5.8kb PstI片段中发现了四个ORF,分别命名为ORF1、ORF2、ORF3和ORF4,该片段先前由Yoshimura、Takahashi、Hibi、Takasawa、Kato和Dickinson克隆并进行了部分特征分析(《感染与免疫》61:5181-5189,1993年)。所有ORF的转录方向与fimA相同。据报道,分别由ORF2和ORF3编码的50 kDa和80 kDa蛋白质是与菌毛相关的次要成分。分别由ORF1和ORF4编码的15 kDa和19 kDa蛋白质已在大肠杆菌中表达,但在牙龈卟啉单胞菌中未鉴定到。然而,基于其N端氨基酸序列,在大肠杆菌中表达的所有ORF基因产物似乎都含有完整的信号肽。