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Microarray-based fluorescence assay of endonuclease functionality and inhibition.
MedLine Citation:
PMID:  23282850     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Here, a double-strand (ds) DNA microarray-based fluorescence assay has been deployed for studying endonuclease functionality and inhibition. The dsDNA microarrays are fabricated by hybridization of the Cy5-labeled oligonucleotides with the immobilized complementary oligonucleotide probes on the glass slide. The microarray displays significant fluorescence decrease in response to endonuclease, which is able to cleave the oligonucleotide moiety of dsDNA. Two endonucleases, EcoRI and BamHI, and four potential inhibitors, doxorubicin hydrochloride (DOX), 5-fluorouracil (5-FU), ethidium bromide (EB) and actinomycin D (ACTD), were selected to address the feasibility of this assay. Enzyme activities of the endonucleases are detected with high specificity down to the limits of 1.1 U mL(-1) for EcoRI and 2.0 U mL(-1) for BamHI, respectively. In addition, BamHI and EcoRI inhibition by the inhibitors are also shown, demonstrating the potential for high-throughput screening for inhibitors.
Authors:
Lan Ma; Min Su; Tao Li; Zhenxin Wang
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-1-3
Journal Detail:
Title:  The Analyst     Volume:  -     ISSN:  1364-5528     ISO Abbreviation:  Analyst     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-1-3     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372652     Medline TA:  Analyst     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, P. R. China. wangzx@ciac.jl.cn.
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