铜绿假单胞菌抗原基因细胞定位分析

Analysis of the cellular location of Pseudomonas aeruginosa antigen genes

目的筛选刺激机体产生抗体的铜绿假单胞菌优势抗原表位,分析铜绿假单胞菌抗原基因的定位。方法收集健康人和铜绿假单胞菌感染患者血清,应用噬菌体展示十二肽库进行1轮阴性选择淘选和3轮生物淘选,对阳性克隆单噬菌体进行测序,与PAO1基因组序列比对分析。结果 30个阳性克隆单噬菌体,测序得到12种序列。与PAO1基因组序列比对得到673个基因,细胞定位为胞浆、内膜、周质、外膜、胞外和未知,所占比例分别为49%、22%、6%、6%、2%和15%。基因产物按功能分为26类,各部位的基因与细胞功能一致。结论推测胞外和外膜中"细胞壁/脂多糖/荚膜"类、"分泌因子"类和"运动及黏附"类抗原可用于疫苗研究。

Objective To identify epitopes of Pseudomonas aeruginosa dominant antigens that can stimulate the body to produce antibodies and analyze the location of antigen genes. Methods The sera collected from controls and patients infected by Pseudomonas aeruginosa were used to screen the immunodominant of mimic epitopes of PA by a random 12-peptide phage display through one negative selection panning and three rounds biopanning. The positive phages clones were sequenced and then compared with the published sequence of strain PAO1. Results After four rounds of biopanning, a significant enrichment of phages were achieved. The inserts fron 30 positive phages clones were devided into 12 peptides based on different amino acids sequence. Through comparison with PAO1 genomic sequences, we got 673 genes, whose cellular localization were cytoplasm, cytoplasmic membrane, periplasm, outer membrane, extracellular and unknown, and the proportions were 49%, 22%, 6%, 6%, 2%and 15%respectively. Gene products could be classified into 26 categories, and the genes in each part of the cells were in accordance with its function. Conclusion We speculated that the antigens of＆quot;cell wall/LPS/capsule＆quot;,＆quot;secreted factors＆quot;and＆quot;motility and attachment＆quot;could be used for vaccine research.