Capillary electrophoresis-SELEX selection of aptamers with affinity for HIV-1 reverse transcriptase
Capillary electrophoresis-SELEX selection of aptamers with affinity for HIV-1 reverse transcriptase
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DOI：
10.1021/ac050836q
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发表时间：
2005-10-01
影响因子：
7.4
通讯作者：
Bowser, MT
中科院分区：
文献类型：
--
作者：
Mosing, RK;Mendonsa, SD;Bowser, MT
采用毛细管电泳selex （CE-SELEX）筛选具有HIV逆转录酶（HIVRT）亲和力的ssDNA适配体。将ssDNA文库与HIVRT孵育。结合HIVRT的序列用CE分离、PCR扩增和纯化，得到一个丰富的ssDNA库，适合进一步的选择。经过4轮筛选，获得解离常数低至180pm的适体。这是首次报道用CE-SELEX分离的适配体比使用常规选择技术获得的适配体具有更高的亲和力。测序的27个克隆中没有发现序列基序，这表明有许多序列可以结合低皮摩尔解离常数的HIVRT。
Capillary electrophoresis-SELEX (CE-SELEX) was used to select ssDNA aptamers with affinity for HIV reverse transcriptase (HIVRT). A library of ssDNA was incubated with HIVRT. Sequences bound to HIVRT were isolated using CE, PCR amplified, and purified, yielding an enriched ssDNA pool suitable for further rounds of selection. Aptamers with dissociation constants as low as 180 pM were isolated after four rounds of selection. This is the first report of aptamers isolated by CE-SELEX with higher affinity than those obtained for the same target using conventional selection techniques. No sequence motifs were identified in the 27 clones sequenced, suggesting that there are many sequences that can bind HIVRT with low picomolar dissociation constants.