We demonstrate the application of superlocalization microscopy to identify sequence-specific portions of single-stranded DNA (ssDNA) with sequence resolution of 50 nucleotides, corresponding to a spatial resolution of 30 nm. Super-resolution imaging was achieved using a variation of a single-molecule localization method, termed as "motion blur" point accumulation for imaging in nanoscale topography (mbPAINT). The target ssDNA molecules were immobilized on the substrate. Short, dye-labeled, and complementary ssDNA molecules stochastically bound to the target ssDNA, with repeated binding events allowing super-resolution. Sequence specificity was demonstrated via the use of a control, noncomplementary probe. The results support the possibility of employing relatively inexpensive short ssDNAs to identify gene sequence specificity with improved resolution in comparison to the existing methods.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934010 | PMC |
| http://dx.doi.org/10.1021/am403984k | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Communication between DNA polymerases and Replication Protein A within the archaeal replisome.
Nat Commun
December 2024
Architecture and Dynamics of Biological Macromolecules, Institut Pasteur, Université Paris Cité, CNRS UMR 3528, Paris, France.
Replication Protein A (RPA) plays a pivotal role in DNA replication by coating and protecting exposed single-stranded DNA, and acting as a molecular hub that recruits additional replication factors. We demonstrate that archaeal RPA hosts a winged-helix domain (WH) that interacts with two key actors of the replisome: the DNA primase (PriSL) and the replicative DNA polymerase (PolD). Using an integrative structural biology approach, combining nuclear magnetic resonance, X-ray crystallography and cryo-electron microscopy, we unveil how RPA interacts with PriSL and PolD through two distinct surfaces of the WH domain: an evolutionarily conserved interface and a novel binding site.
View Article and Find Full Text PDFPyDNA-templated AgNPs coupled with poly (β‑cyclodextrin) enhanced fluorescence: A facile platform for signal amplification detection of biothiols in living cells and zebrafish.
Spectrochim Acta A Mol Biomol Spectrosc
December 2024
School of Pharmacy, Xinxiang Medical University, Xinxiang, Henan 453003, PR China.
Accurate and sensitive fluorescence imaging of biothiols is essential for understanding the mechanism underlying some physiological and pathological events, as well as the prevention and diagnosis of diseases. However, low signal transduction efficiency and poor biocompatibility of fluorescence tags associated with current sensors hinder their potential utilizations. Herein, a smart biothiols sensitive vivo imaging platform on the basis of amplifying nanoprobe has been designed.
View Article and Find Full Text PDFLEF3 phosphorylation attenuates the replication of Bombyx mori nucleopolyhedrovirus by suppressing its interaction with alkaline nuclease.
Virology
December 2024
Institute of Biochemistry, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Zhejiang Province, 310018, Hangzhou, China; Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine, Zhejiang Province, 310018, Hangzhou, China. Electronic address:
Late expression factor 3 (LEF3), a multifunctional single-stranded DNA binding protein encoded by baculoviruses, is indispensable for viral DNA replication and plays a pivotal role in viral infection. Our previous quantitative analysis of phosphorylomics revealed that the phosphorylation levels of two serine residues (S8 and S25) located in LEF3 nuclear localization sequence were significantly up-regulated after Bombyx mori nucleopolyhedrovirus (BmNPV) infection, but the underlying mechanism remained unknown. To investigate the impact of phosphorylation on BmNPV infection, site-direct mutagenesis was performed on LEF3 to obtain phosphorylated mimic (S/D) or dephosphorylated mimic (S/A) mutants.
View Article and Find Full Text PDFFour SNPs in the untranslated region of the NOTCH2 gene involved in reproductive traits in fertile and anestrum Holstein Friesian cattle.
BMC Vet Res
December 2024
Department of Research, Research and Development Station for Bovine, Arad, Romania.
Background: There are no studies belong NOTCH2 gene polymorphism in relation to reproductive and productive traits in Holstein cattle. The objective of the present study was to investigate the effect of NOTCH2 gene polymorphisms on productive and reproductive performance of fertile and anestrum cattle.
Methods: The cattle were classified into anestrus for 3-12 months postpartum (n = 115, 37.
Anti-miR21-conjugated DNA nanohydrogel for enhanced cancer therapy.
Biomater Adv
December 2024
Department of Chemistry and the Natural Science Research Institute, Myongji University, 116 Myongji-ro, Yongin-si 17058, Republic of Korea. Electronic address:
MicroRNAs (miRNAs) are non-coding, endogenous small single-stranded RNA molecules involved in post-transcriptional regulation of gene expression. It has been demonstrated that dysregulation of miRNA plays a major role in tumor formation, proliferation, and metastasis. Therefore, the delivery of anti-miRNA oligonucleotides to block the activity of these oncogenic miRNAs is a high-potential anti-cancer therapy approach.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!