DNA single-base mismatch study with an electrochemical enzymatic genosensor.
DNA single-base mismatch study with an electrochemical enzymatic genosensor.
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DOI：
10.1016/j.bios.2006.07.015
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发表时间：
2007-03-15
影响因子：
12.6
通讯作者：
Costa-García A
中科院分区：
文献类型：
--
作者：
Abad-Valle P;Fernández-Abedul MT;Costa-García A
采用电化学酶基因传感器的DNA杂交的一个彻底的选择性研究进行了讨论。将30-mer 3′-巯基化DNA链固定在金膜上后，与生物素化互补DNA链发生杂交。然后，碱性磷酸酶通过链霉亲和素-生物素相互作用掺入双链体。从3-吲哚酚磷酸和随后的电化学检测进行了酶促生成靛蓝。研究了杂交条件的影响，以更好地区分完全互补和错配的链。可以检测3、2和1个错配。研究了单碱基错配的类型和位置，以及链长对错配的影响。也考虑假定阅读框移位的突变。测试了浓度对选择性的影响，得到具有足够灵敏度和稳定性的高选择性基因传感器。
A thorough selectivity study of DNA hybridization employing an electrochemical enzymatic genosensor is discussed here. After immobilizing on a gold film a 30-mer 3′-thiolated DNA strand, hybridization with a biotinylated complementary one takes place. Then, alkaline phosphatase is incorporated to the duplex through the interaction streptavidin–biotin. Enzymatic generation of indigo blue from 3-indoxyl phosphate and subsequent electrochemical detection was made. The influence of hybridization conditions was studied in order to better discern between fully complementary and mismatched strands. Detection of 3, 2 and 1 mismatch was possible. The type and location of the single-base mismatch, as well as the influence of the length of the strands was studied too. Mutations that suppose displacement of the reading frame were also considered. The effect of the concentration on the selectivity was tested, resulting a highly selective genosensor with an adequate sensitivity and stability.
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DOI：
10.1073/pnas.071623998
发表时间：
2001-03-27
影响因子：
11.1
作者：
Heaton, RJ;Peterson, AW;Georgiadis, RM
通讯作者：
Georgiadis, RM
影响因子：
4.5
作者：
Campuzano, S;Gálvez, R;Pingarrón, JM
通讯作者：
Pingarrón, JM
影响因子：
14.9
作者：
IKUTA, S;TAKAGI, K;ITAKURA, K
通讯作者：
ITAKURA, K
影响因子：
2.9
作者：
Allawi, HT;SantaLucia, J
通讯作者：
SantaLucia, J
影响因子：
2.9
作者：
Allawi, HT;SantaLucia, J
通讯作者：
SantaLucia, J