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用SELEX技术 ,寻获炭疽芽孢适配子 ,研究其亲和功能及是否作为炭疽芽孢的检测分子。化学合成长度为 35mer的随机DNA库 ,以炭疽杆菌疫苗株A .16R芽孢为靶标进行 18轮的筛选 ,筛选产物克隆、测序 ,利用生物素 亲和素 辣根过氧化物酶显色系统判断适配子与芽孢的结合活性 ;计算机软件分析测序适配子保守序列和二级结构 ;以兔抗炭疽芽孢抗体为捕获分子 ,适配子为检测分子混合夹心法检测炭疽芽孢。第 18轮筛选的适配子与芽孢结合后A值比第 1轮的提高了 3733 .33 %以上 ;测序 79个序列中 ,A值最高为 1.2 0 ,最低为 0 .2 0 ;混合夹心法检测表明 ,适配子量为 16 μg ,芽孢为 4× 10 7个时 ,显色信号强度最强。结果提示 ,其 5′端茎环及发夹结构是与芽孢结合的基础 ,远程高级结构对其结合功能有一定的影响 ;寡核苷酸适配子可以作为炭疽芽孢的检测分子
SELEX technology was used to find anthrax spore aptamers to study their affinity function and whether they were used as detection molecules for anthrax spores. Chemical synthesis of a random length of 35mer DNA library to Bacillus anthracis vaccine strain A. 16R spores as the target for 18 rounds of screening, screening products, sequencing, the use of biotin-avidin horseradish peroxidase color system to determine the appropriate Gametophyte and spore binding activity; computer software sequencing sequence analysis aptamer conserved sequence and secondary structure; rabbit anti-anthrax spores antibodies capture molecules, aptamers for the detection of molecular mixture sandwich method for the detection of anthrax spores. The A value of the 18th screening aptamers combined with spores was higher than that of the first round by more than 3733.33%. Among the 79 sequences sequenced, the highest A value was 1.2 0 and the lowest was 0. 2, The results showed that when the amount of aptamer was 16 μg and the number of spores was 4 × 10 7, the color signal intensity was the strongest. The results suggest that the 5 ’end of the stem ring and hairpin structure is the basis of the combination with spores, the remote high-level structure of its binding function to some extent; oligonucleotide aptamers can be used as a detection of anthrax spores molecules