TY - JOUR
T1 - 闭锁连接蛋白-1 核酸适配体的筛选与初步应用
AU - Li, Yujuan
AU - Zhang, Hanwen
AU - Liu, Shan
AU - Wang, Jiaping
AU - Li, Yongzhi
N1 - Publisher Copyright:
© 2023 Beijing Institute of Technology. All rights reserved.
PY - 2023/8
Y1 - 2023/8
N2 - To select the Aptamer (APT) of Zonula Occludens-1 (ZO-1), a systematic evolution technique of ligands was used based on exponential enrichment (SELEX), and 15 APTZO-1 were selected as Aptamers finally. Excluding linear and similar structures, an enzyme-linked immune-sorbent assay was used to evaluate the affinity between 9 APTsZO-1 and ZO-1. The analysis results show the order of affinity, APTZO-1 8 > APTZO-1 2 > APTZO-1 7 > APTZO-1 13 > APTZO-1 1 > APTZO-1 5 > APTZO-1 15 > APTZO-1 12 > APTZO-1 4, signed with Cy5 fluorescent labeling, for instance, Cy5-APTZO-1 8 labeling for APTZO-1 8. The enzyme-linked immune-sorbent method can be used for fluorescence localization and semi-quantitative analysis of ZO-1 for human epithelial colorectal adenocarcinoma cells (Caco-2) and human brain capillary endothelial cells (hCMEC/D3). And the result is identical with results of immunofluorescence and Western blotting based on ZO-1 antibody methods. In the study, a series of APTs for ZO-1 are firstly screened and expected to provide a new tool molecule with high specificity for fluorescent labeling and semi-quantitative analysis of ZO-1.
AB - To select the Aptamer (APT) of Zonula Occludens-1 (ZO-1), a systematic evolution technique of ligands was used based on exponential enrichment (SELEX), and 15 APTZO-1 were selected as Aptamers finally. Excluding linear and similar structures, an enzyme-linked immune-sorbent assay was used to evaluate the affinity between 9 APTsZO-1 and ZO-1. The analysis results show the order of affinity, APTZO-1 8 > APTZO-1 2 > APTZO-1 7 > APTZO-1 13 > APTZO-1 1 > APTZO-1 5 > APTZO-1 15 > APTZO-1 12 > APTZO-1 4, signed with Cy5 fluorescent labeling, for instance, Cy5-APTZO-1 8 labeling for APTZO-1 8. The enzyme-linked immune-sorbent method can be used for fluorescence localization and semi-quantitative analysis of ZO-1 for human epithelial colorectal adenocarcinoma cells (Caco-2) and human brain capillary endothelial cells (hCMEC/D3). And the result is identical with results of immunofluorescence and Western blotting based on ZO-1 antibody methods. In the study, a series of APTs for ZO-1 are firstly screened and expected to provide a new tool molecule with high specificity for fluorescent labeling and semi-quantitative analysis of ZO-1.
KW - Aptamer (APT)
KW - Zonula Occludens-1(ZO-1)
KW - fluorescent labeling
KW - semi-quantitative analysis
KW - systematic evolution of ligands by exponential enrichment(SELEX)
UR - http://www.scopus.com/inward/record.url?scp=85170232396&partnerID=8YFLogxK
U2 - 10.15918/j.tbit1001-0645.2022.226
DO - 10.15918/j.tbit1001-0645.2022.226
M3 - 文章
AN - SCOPUS:85170232396
SN - 1001-0645
VL - 43
SP - 870
EP - 877
JO - Beijing Ligong Daxue Xuebao/Transaction of Beijing Institute of Technology
JF - Beijing Ligong Daxue Xuebao/Transaction of Beijing Institute of Technology
IS - 8
ER -