Screening and characterization of aptamers of human chronic myeloid leukemia (CML) fusion protein BCR-ABL and its structure analysis
10.3969/j.issn.1000-484X.2015.01.017
- VernacularTitle:人慢性粒细胞白血病融合蛋白BCR－ABL适配子筛选及结构分析
- Author:
Juan PING
;
Na ZHAO
;
Zhihui SHEN
;
Mingxing YIN
;
Qian ZHANG
;
Wei ZHANG
;
Xueshan MA
;
Chuanbo CHEN
- Publication Type:Journal Article
- Keywords:
SELEX technology;
Human chronic myeloid leukemia;
BCR-ABL fusion protein;
Aptamers;
Screen
- From:
Chinese Journal of Immunology
2015;(1):82-85
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To screen and characterize aptamers against BCR-ABL fusion protein.Methods:A 90bp single stranded DNA( ssDNA) random library was subjected to 13 rounds of selection against BCR-ABL fusion protein by systematic evolution of ligands by expotential enrichment ( SELEX ) method, the selected aptamers were cloned and sequenced.The primary sequences and structure of aptamers were analyzed by Clustal W and DNA Folding Sever and the percentage of the ssDNA pool bound to BCR-ABL core protein were determinated.Results: after 13 rounds selection, the percentage of ssDNA pool bound to BCR-ABL fusion protein increased from 0.3%to 47.1%,the results showed that affinities of the Aptamers were different,the second structure analysis revealed possible stem-loops for binding to BCR-ABL fusion protein,the affinity of aptamer A2 to BCR-ABL fusion protein was highest with Kd values as low as 72 nmol/L.Conclusion:Aptamers against BCR-ABL fusion protein has been identified by SELEX methods from a 90 bp single stranded DNA library.And provide certain reference for the clinical treatment of chronic myelogenous.
