期刊论文

中文

北京大学学报(医学版)

1671-167X

2014

46

5

698-702

10.3969/j.issn.1671-167X.2014.05.006

(81202451,81000026):国家自然科学基金 (12JJ4029):湖南省自然科学基金 (20114323120002):高等学校博士学科点专项科研基金 (2012M511731):中国博士后基金 (11A083):湖南省教育厅科学研究项目重点项目 :湖南省高校科技创新团队基金(15) (100601):湖南省重点学科基金 (81000026 and 81202451):Supported by the National Natural Sciences Foundation of China (12JJ4029):the National Natural Science Foundation of Hunan,China (20114323120002):the Special Scientific Research Foundation for Doctoral Discipline Area of the Institution of Higher Learning of China (2012M511731):the National Science Foundation for Post-Doctoral Scientists of China (11 A083):Research Foundation of Education Bureau of Hunan Province,China :Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province(15) (100601):the Construct Program of the Key Discipline In Hunan Province

本校为第一机构

目的:筛选大肠埃希菌外排泵tolC蛋白的核酸适配体。方法:重组表达大肠埃希菌外排泵外膜蛋白tolC,利用指数富集的配体系统进化技术(stematic evolution of ligand by exponential enrichment,SELEX)从单链DNA文库中筛选出一组能够特异性与其结合的核酸适配体。利用FITC荧光素标记技术检测每轮单链DNA文库与靶蛋白的亲和力,直至亲和力的上升趋于饱和,将最后一轮筛选产物克隆测序,并用DNAman软件分析其二级结构。结果:经过12轮筛选,单链DNA文库与靶标蛋白的亲和力趋向稳定,将第12轮筛选产物克隆测序,对获得的23个适配体进行分析。一级结构分析显示23个适配体并无共同的保守序列,但分别有3对和2对适...

OBJECTIVE: To screen and characterize the aptamer of Escherichia coli outer member protein tolC. METHODS: By using the recombinant E.coli outer member protein tolC for the screening target, oligonucleotides which were capable of specifically binding to the protein were screened from a random oligonucleotide library through the stematic evolution of ligand by exponential enrichment (SELEX) technique. The binding capacity of ssDNA to the targeted protein from each round was detected by the FITC fluorescence labeling technique.The ssDNA from the last cycle was cloned and sequenced,and the second ...