Single cell-level detection and quantitation of leaky protein expression from any strongly regulated bacterial system
2015
Arora, Kanika | Mangale, Sachin S. | Guptasarma, Purnananda
Extremely low levels of “leaky” expression of genes in bacterial protein expression systems can severely curtail cell viability when expressed proteins are toxic. A general method for sensitive detection of such expression is lacking. Here, we present a method based on microscopic visualization of a fluorescent “reporter” protein (RFP–HU-A) constructed by fusing red fluorescent protein (RFP) to the N-terminus of a nucleoid-associated, histone-like DNA-binding protein, HU-A. Localization of RFP–HU-A within nucleoids facilitates detection, quantitation, and characterization of leaky expression at the single-cell level.
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Эту запись предоставил National Agricultural Library