Burucoa C, Frémaux C, Pei Z, Tummuru M, Blaser M J, Cenatiempo Y, Fauchère J L
Laboratoire de Microbiologie A, CHU Poitiers, France.
Res Microbiol. 1995 Jul-Aug;146(6):467-76. doi: 10.1016/0923-2508(96)80292-0.
The 29-kDa protein PEB4, a major antigen of Campylobacter jejuni, is present in all C. jejuni strains tested and elicits an antibody response in infected patients. By screening a lambda gt11 library of chromosomal DNA fragments of C. jejuni strain 81-176 in Escherichia coli Y1090 cells with antibody raised against purified PEB4, a recombinant phage with a 2-kb insert expressing an immunoreactive protein of 29 kDa was isolated. DNA sequence analysis revealed that the insert contains two complete open reading frames ORF-A and ORF-B. ORF-A (peb4A) encodes a 273-residue protein with a calculated molecular mass of 30,460 daltons. The deduced amino acid sequence, composition and pl of the recombinant mature protein are similar to those determined for purified PEB4. The first 21 residues resemble a signal peptide. Gene bank searches indicated 33.7% identity with protein export protein PrsA of Bacillus subtilis and 23.8% identity with protease maturation protein precursor PrtM of Lactococcus lactis. PCR experiments indicate that peb4A is highly conserved among C. jejuni strains. ORF-B begins 2 bp after the last codon of peb4A and encodes a putative protein of 353 residues with 63.4% identity with E. coli fructose 1,6-biphosphate aldolase. The sequence arrangement suggests that these two genes form an operon.
29 kDa的蛋白质PEB4是空肠弯曲菌的主要抗原，存在于所有检测的空肠弯曲菌菌株中，并能在感染患者体内引发抗体反应。用针对纯化的PEB4产生的抗体筛选空肠弯曲菌81 - 176株染色体DNA片段的λgt11文库，在大肠杆菌Y1090细胞中分离出一个带有2 kb插入片段、表达29 kDa免疫反应性蛋白的重组噬菌体。DNA序列分析表明，该插入片段包含两个完整的开放阅读框ORF - A和ORF - B。ORF - A（peb4A）编码一个273个残基的蛋白质，计算分子量为30,460道尔顿。推导的重组成熟蛋白的氨基酸序列、组成和等电点与纯化的PEB4所测定的相似。前21个残基类似于信号肽。基因库搜索表明，它与枯草芽孢杆菌的蛋白质输出蛋白PrsA有33.7%的同一性，与乳酸乳球菌的蛋白酶成熟蛋白前体PrtM有23.8%的同一性。PCR实验表明，peb4A在空肠弯曲菌菌株中高度保守。ORF - B在peb4A的最后一个密码子后2 bp处开始，并编码一个353个残基的推定蛋白，与大肠杆菌果糖1,6 - 二磷酸醛缩酶有63.4%的同一性。序列排列表明这两个基因形成一个操纵子。
