TITLE:
High yield expression of proteins in E. coli for NMR studies
AUTHORS:
Somnath Mondal, Divya Shet, Chinmayi Prasanna, Hanudatta S. Atreya
KEYWORDS:
E. Coli; Recombinant DNA Technology; Structural Biology; NMR Spectroscopy
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.4 No.6,
June
24,
2013
ABSTRACT:
In recent years, high yield expression of proteins in E.
coli has witnessed rapid progress with developments of new methodologies
and technologies. An important advancement has been the development of novel recombinant cloning approaches and protocols to express heterologous proteins
for Nuclear Magnetic Resonance (NMR) studies and for isotopic enrichment.
Isotope labeling in NMR is necessary for rapid acquisition of high dimensional
spectra for structural studies. In addition, higher yield of proteins using
various solubility and affinity tags has made protein over-expression
cost-effective. Taken together, these methods have opened new avenues for
structural studies of proteins and their interactions. This article deals
with the different techniques that are employed for over-expression of proteins
in E. coli and different methods used for isotope labeling of proteins
vis-à-vis NMR spectroscopy.