Bacteriophage strain typing by rapid single molecule analysis
Artikel i vetenskaplig tidskrift, 2015

Rapid characterization of unknown biological samples is under the focus of many current studies. Here we report a method for screening of biological samples by optical mapping of their DNA. We use a novel, one-step chemo-enzymatic reaction to covalently bind fluorophores to DNA at the four-base recognition sites of a DNA methyltransferase. Due to the diffraction limit of light, the dense distribution of labels results in a continuous fluorescent signal along the DNA. The amplitude modulations (AM) of the fluorescence intensity along the stretched DNA molecules exhibit a unique molecular fingerprint that can be used for identification. We show that this labelling scheme is highly informative, allowing accurate genotyping. We demonstrate the method by labelling the genomes of lambda and T7 bacteriophages, resulting in a consistent, unique AM profile for each genome. These profiles are also successfully used for identification of the phages from a background phage library. Our method may provide a facile route for screening and typing of various organisms and has potential applications in metagenomics studies of various ecosystems.

Författare

A. Grunwald

Tel Aviv University

M. Dahan

Tel Aviv University

A. Giesbertz

RWTH Aachen University

A. Nilsson

Lunds universitet

Lena Nyberg

Chalmers, Biologi och bioteknik, Kemisk biologi

E. Weinhold

RWTH Aachen University

T. Ambjornsson

Lunds universitet

Fredrik Westerlund

Chalmers, Biologi och bioteknik, Kemisk biologi

Y. Ebenstein

Tel Aviv University

Nucleic Acids Research

0305-1048 (ISSN) 1362-4962 (eISSN)

Vol. 43 18 e117- e117

Ämneskategorier

Biokemi och molekylärbiologi

Styrkeområden

Livsvetenskaper och teknik

DOI

10.1093/nar/gkv563

Mer information

Senast uppdaterat

2018-03-27