Duffy Michael F, Walker Ian D, Browning Glenn F
Faculty of Veterinary Science, Veterinary Preclinical Centre, The University of Melbourne, Parkville, Victoria, Australia 3052.
Microbiology (Reading). 1997 Oct;143 ( Pt 10):3391-3402. doi: 10.1099/00221287-143-10-3391.
Sera from 10 patients infected with Mycoplasma pneumoniae were used in Western blot analysis of Triton-X-114-soluble protein preparations of M. pneumoniae. All 10 sera were reactive with a protein antigen of 116 kDa. Sera from another 17 patients were used in Western blot analysis of whole-cell M. pneumoniae proteins; 15 of these sera were reactive with the 116 kDa protein. Trypsin digestion of whole M. pneumoniae cells demonstrated the surface location of this protein. Sequencing of DNA which contained the gene for this protein identified an ORF of 3093 bp encoding a protein with a predicted molecular mass of 116013 Da. The ORF for the 116 kDa protein had 99.8% nucleotide identity with the M. pneumoniae gene G07_orf1030 and 61% nucleotide identity with the Mycoplasma genitalium ORF MG075 of unassigned function. An ORF which was identified 5' to the 116 kDa protein ORF coded for a 16 kDa protein and had 99.8% nucleotide identity with the M. pneumoniae gene G07_orf135 and 58.4% nucleotide identity with the ORF MG074 of M. genitalium. Analysis of mRNA detected a 3.7 kb transcript with a single initiation site 5' to the ORF encoding the 16 kDa protein. The coding sequences for both the 16 kDa protein and the 116 kDa protein were present in this transcript, indicating that they were part of an operon and suggesting a possible functional relationship.
将10例感染肺炎支原体患者的血清用于肺炎支原体Triton-X-114可溶性蛋白制剂的蛋白质印迹分析。所有10份血清均与一种116 kDa的蛋白质抗原发生反应。另外17例患者的血清用于肺炎支原体全细胞蛋白的蛋白质印迹分析；其中15份血清与116 kDa蛋白发生反应。肺炎支原体全细胞的胰蛋白酶消化证明了该蛋白的表面定位。对包含该蛋白基因的DNA进行测序，确定了一个3093 bp的开放阅读框，其编码的蛋白预测分子量为116013 Da。116 kDa蛋白的开放阅读框与肺炎支原体基因G07_orf1030的核苷酸同一性为99.8%，与未明确功能的生殖支原体开放阅读框MG075的核苷酸同一性为61%。在116 kDa蛋白开放阅读框5'端发现的一个开放阅读框编码一种16 kDa蛋白，与肺炎支原体基因G_{07}_orf135的核苷酸同一性为99.8%，与生殖支原体开放阅读框MG074的核苷酸同一性为58.4%。对mRNA的分析检测到一个3.7 kb的转录本，其在编码16 kDa蛋白的开放阅读框5'端有一个单一的起始位点。16 kDa蛋白和116 kDa蛋白的编码序列均存在于该转录本中，表明它们是一个操纵子的一部分，并提示可能存在功能关系。
