Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:6.5.1.2 (
DNA ligase
)
2,749
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
RNA modification, such as
N
1-methyladenosine (m
1
A), affects the secondary structure of RNA and its ability to recognize specific reader proteins. Methods for detecting site-specific m
1
A are in demand. We report here a ligation-assisted differentiation approach for quantitative detection of m
1
A in mRNA with single-base resolution. The methyl group in m
1
A disrupts the Watson-Crick base pairing with uridine, resulting in a lower ligation efficiency of certain ligases and lower amounts of ligation products. Detection of the ligation products using quantitative real-time PCR provided site-specific evaluation of m
1
A. We first screened appropriate ligase and found that T3
DNA ligase
offered the best discrimination between m
1
A and adenosine. We successfully detected and quantified m
1
A at position 1674 of bromodomain containing 2 (
BRD2
) mRNA from HEK293T cells. In lung carcinoma tissues, the level of m
1
A at position 1674 of
BRD2
mRNA was significantly decreased compared to the tumor-adjacent normal tissues, suggesting that site-specific m
1
A may be involved in carcinogenesis.
...
PMID:Quantification and Single-Base Resolution Analysis of
N
1-Methyladenosine in mRNA by Ligation-Assisted Differentiation. 3194 30
