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目的 :利用SELEX(systemevolutionofligandsbyexponentialenrichment)体外筛选技术 ,寻找能与炭疽芽孢杆菌芽孢特异结合的寡核苷酸适配子 (aptamer)。方法 :体外合成长度为 78个核苷酸的随机DNA库 ,通过SELEX技术 ,以炭疽芽孢杆菌疫苗株A .16R的芽孢为靶标进行 15轮的筛选 ,利用生物素 亲和素显色系统判断寡核苷酸与芽孢的结合活性。结果 :随着筛选轮数的增加 ,PCR扩增电泳条带逐渐单纯 ,寡核苷酸与芽孢结合后显色 ,D值提高了 9倍以上 ,D值随适配子结合量的增加而递增。结论 :已初步筛选到与炭疽芽孢具有亲和力的适配子。
OBJECTIVE: To use the SELEX (systemevolutionofligandsbyexponentialenrichment) in vitro screening technique to find oligonucleotide aptamers that specifically bind to B. anthracis spores. Methods: A randomized DNA library with a length of 78 nucleotides was synthesized in vitro. SELEX technology was used to screen 15 spores of Bacillus anthracis vaccine strain A16R for 15 cycles. The biotin-avidin- Nucleotide and spore binding activity. Results: With the increase of the number of rounds of selection, the band of PCR amplification was gradual and simple. The combination of oligonucleotide and spores was more obvious. The D value increased more than 9 times. The value of D increased with the increase of aptamer binding . CONCLUSION: Aptamers with affinity to anthracis are initially screened.