Long-lived intracellular single-molecule fluorescence using electroporated molecules.

ABSTRACT: Studies of biomolecules in vivo are crucial to understand their function in a natural, biological context. One powerful approach involves fusing molecules of interest to fluorescent proteins to study their expression, localization, and action; however, the scope of such studies would be increased considerably by using organic fluorophores, which are smaller and more photostable than their fluorescent protein counterparts. Here, we describe a straightforward, versatile, and high-throughput method to internalize DNA fragments and proteins labeled with organic fluorophores into live Escherichia coli by employing electroporation. We studied the copy numbers, diffusion profiles, and structure of internalized molecules at the single-molecule level in vivo, and were able to extend single-molecule

SUBMITTER: Crawford R 

PROVIDER: S-EPMC3853080 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature