Expression regulation by a methyl-CpG binding domain in an E. coli based, cell-free TX-TL system.

@article{Schenkelberger2017ExpressionRB,
  title={Expression regulation by a methyl-CpG binding domain in an E. coli based, cell-free TX-TL system.},
  author={Marc Schenkelberger and Siba Shanak and Marc Finkler and Emanuel G Worst and Vincent Noireaux and Volkhard Helms and Albrecht Ott},
  journal={Physical biology},
  year={2017},
  volume={14 2},
  pages={
          026002
        }
}
Cytosine methylation plays an important role in the epigenetic regulation of eukaryotic gene expression. The methyl-CpG binding domain (MBD) is common to a family of eukaryotic transcriptional regulators. How MBD, a stretch of about 80 amino acids, recognizes CpGs in a methylation dependent manner, and as a function of sequence, is only partly understood. Here we show, using an Escherichia coli cell-free expression system, that MBD from the human transcriptional regulator MeCP2 performs as a… 
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Methylation-targeted specificity of the DNA binding proteins R.DpnI and MeCP2 studied by molecular dynamics simulations
TLDR
The results of molecular dynamics simulations, alchemical free energy perturbation, and MM-PBSA calculations are presented to explain the specificity of the R.DpnI enzyme from Streptococcus pneumonia in binding to adenine-methylated DNA with both its catalytic and winged-helix domains.
How a single 5-methylation of cytosine regulates the recognition of C/EBPβ transcription factor: a molecular dynamic simulation study
TLDR
Comparisons of the structures and dynamics of CEBP/βprotein–DNA complexes before and after methylation indicate that even a single 5mC can regulate protein–DNA recognition by steric-hindrance effect of methyl group and changing the hydrogen bond interactions.
A Methylation-Directed, Synthetic Pap Switch Based on Self-Complementary Regulatory DNA Reconstituted in an All E. coli Cell-Free Expression System.
TLDR
This work uses an E. coli cell-free expression system to study molecular functions of the pap regulatory region based on a specially designed, synthetic construct flanked by two reporter genes encoding fluorescent proteins for simple readout and suggests that besides Lrp, the conformation of the self-complementary regulatory DNA plays a strong role in the regulation of phase-variation.
DNA methylation and the core pluripotency network.
Full incorporation of the noncanonical amino acid hydroxylysine as a surrogate for lysine in green fluorescent protein.
A bead-based method for the removal of the amino acid lysine from cell-free transcription-translation systems
TLDR
A two-step method for the removal of residual free lysine in an all Escherichia coli-based cell-free expression system is proposed, which is an efficient, fast and simple way to remove residualLysine without altering the system ability to perform gene expression.
A bead-based method for the removal of the amino acid lysine from cell-free transcription-translation systems.
Research Status and Application Prospect of Epigenetic Regulation of BDNF Gene Expression in Chronic Pain

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