| 铜绿假单胞菌XN-1随机重组子文库的构建及其在疫苗抗原筛选中的应用研究 |
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| 引用本文： | 许婉婷,顾江,万闯,程新,杨峰,王颖,雷嫏嬛,王兴勇. 铜绿假单胞菌XN-1随机重组子文库的构建及其在疫苗抗原筛选中的应用研究[J]. 免疫学杂志, 2018, 0(12): 1021-1026,1040 |
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| 作者姓名： | 许婉婷 顾江 万闯 程新 杨峰 王颖 雷嫏嬛 王兴勇 |
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| 作者单位： | 重庆医科大学附属儿童医院PICU;陆军军医大学药学与检验医学系微生物与生化药学教研室;儿童发育疾病研究教育部重点实验室、重庆市儿童发育重大疾病诊治与预防国际科技合作基地、儿科学重庆市重点实验室 |
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| 基金项目： | 国家自然科学基金(81772155). |
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| 摘 要： | 目的构建铜绿假单胞菌(Pseudomonas aeruginosa,PA)XN-1的全基因组文库,筛选铜绿假单胞菌新的疫苗候选抗原。方法提取我国分离的临床PA XN-1菌株全基因组,Sau3AⅠ酶切成随机片段后经连接至Bam HⅠ酶切后的pMal-c5x载体,转化至X-blue获得随机重组子。随机重组子经IPTG诱导后使用溶菌酶裂解,收集含有融合的麦芽糖结合蛋白(MBP)的随机重组子的裂解上清,制成含有PA全基因组的蛋白随机文库。通过ELISA方法分别检测单个随机文库蛋白的免疫反应性。将经典的PA强反应抗原PcrV做为参照,分别计算单个随机抗原的相对反应强度并进行比较。提取反应较强的重组子的质粒,测定DNA序列后进行翻译和比对,得到整个强反应性抗原的氨基酸信息。结果成功构建PA XN-1的全基因组文库,筛选到强反应性抗原13个,其中4个抗原的反应强度超过PcrV,分别为Peptidyl-prolyl cis-trans isomerase D、Phosphoenolpyruvate-protein phosphotransferase(PtsP)、Phosphate-starvation-inducible E和Very short patch repair endonuclease。结论基于高效表达系统的PA全基因组文库构建成功,可用于PA及其它病原的疫苗候选抗原的筛选研究。
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| 关 键 词： | 铜绿假单胞菌 全基因组文库 抗原筛选 |
Construction of a random recombinants library of Pseudomonas aeruginosa XN-1 and its application in screening of vaccine candidates |
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| XU Wanting,GU Jiang,WAN Chuang,CHENG Xin,YANG Feng,WANG Ying,LEI Langhuan,WANG Xingyong. Construction of a random recombinants library of Pseudomonas aeruginosa XN-1 and its application in screening of vaccine candidates[J]. Immunological Journal, 2018, 0(12): 1021-1026,1040 |
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| Authors: | XU Wanting GU Jiang WAN Chuang CHENG Xin YANG Feng WANG Ying LEI Langhuan WANG Xingyong |
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| Affiliation: | (Department of Pediatric Intensive Care Unit,Children's Hospital of Chongqing Medical University,Chongqing 400014,China;Department of Microbial and Biochemical Pharmacy,College of Pharmacy and Laboratory Medicine,Army Medical University,Chongqing 400038,China;Ministry of Education Key Laboratory of Child Development and Disorders/China International Science and Technology Cooperation Base of Child Development and Critical Disorders/Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China) |
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| Abstract: | XU Wanting;GU Jiang;WAN Chuang;CHENG Xin;YANG Feng;WANG Ying;LEI Langhuan;WANG Xingyong(Department of Pediatric Intensive Care Unit,Children's Hospital of Chongqing Medical University,Chongqing 400014,China;Department of Microbial and Biochemical Pharmacy,College of Pharmacy and Laboratory Medicine,Army Medical University,Chongqing 400038,China;Ministry of Education Key Laboratory of Child Development and Disorders/China International Science and Technology Cooperation Base of Child Development and Critical Disorders/Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China) |
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| Keywords: | Pseudomonas aeruginosa Antigen library Screening |
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