丙型肝炎病毒非结构蛋白NS2核酸适配子的筛选  

Isolation of DNA aptamers specific for NS2 protein of HCV

作　　者：徐国胜[1] 黄斯瑜[2] 戎霞[1] 王敏[1] 黄可君[1] XU Guo-sheng HUANG Si-yu RONG Xia HUANG Min HUANG Ke-jun(Guangzhou Blood Center, Guangzhou , Guangdong 510095 Zengcheng Blood Station, Guangzhou , Guangdong 511300, China)

机构地区：[1]广州血液中心,广东广州51009 [2]广州血液中心增城区血站,广东广州511300

出　　处：《热带医学杂志》2017年第7期880-883,962,共5页Journal of Tropical Medicine

基　　金：广东省医学科研基金(A2015538)

摘　　要：目的获得与丙型肝炎病毒非结构蛋白NS2特异结合的高亲和力的DNA适配子。方法应用配基指数富集的系统进化(SELEX)技术进行HCV-NS2蛋白适配子筛选,将筛选得到的适配子克隆测序,应用DNAMAN软件对适配子一级结构和二级结构进行比对分析。结果重复筛选7轮后,适配子亲和力和特异性都达到最高,克隆测序得到4个适配子序列,命名为A1、A2、A3、A4,其随机序列部分分别为A1:GTGCGTCCCATGCTGCTGACTTAAATGGGTGGAGGGCAG,A2:CGTGAAATTGTTGACCACTCATGGAATCTGATCTCGTTT,A3:GAAAGGGGATAATCACTTAGGCCTCTCGAATAGTTTATC,A4:AGAAAGTTGAGAACTGCTGTTATTTTGTTAACGTACATG。经软件分析,适配子之间没有共同保守序列和同源序列,适配子的二级空间结构以茎环和口袋结构为主。结论获得了能与HCVNS2蛋白高亲和力和特异性结合的DNA适配子。Objective To find small DNA molecules that are specific and high-affinity ligands of nonstructural 2 （NS2） of HCV and explore the primary structure and secondary structure of the DNA aptamers. Methods Specific and high-affinity aptamers of HCV-NS2 protein were screened by using technology of the Sytematic Evolution of Ligands by Expo-nential enrichment（SELEX） and the structure of the aptamers were analyzed by the DNAMAN software. Results After 7 rounds of selection, 4 NS2 specific aptamers were obtained and named as A1, A2, A3, A4. The random sequences of these aptamers were listed below： A1：GTGCGTCCCATGCTGCTGACTTAAATGGGTGGAGGGCAG A2：CGTGAAATTGTTGACCACTCATGGAA TCTGATCTCGTTT A3：GAAAGGGGATAATCACTTAGGCCTCTCGAATAGTTTATC A4：AGAAAGTTGAGAACTGCTGTTAT TTTGTTAACGTACATG. There were no conservative sequence and homologous sequence among these aptamers. The main secondary structure of these aptamers was stem-loop structure and pocket structure. Conclusion The aptamers were specific and could bind to HCV-NS2 protein with high-affinity.

关 键 词：SELEX 丙型肝炎病毒 NS2蛋白 适配子 

分 类 号：R394.3[医药卫生—医学遗传学]