User:Etienne Robillard/Notebook/Agent Ecoli:PhosphohistidineTransferaseComponent
Agent Ecoli:Phosphohistidine Transferase Compound
OmpR and LuxR multi-step catalytic gene repression systems: Detection and forensic characterization of recombinant (synthetic) DNA material in vivo using dual-input, reversible enzymatic promoters
- Reversion and repression of low to medium copy synthetic DNA plasmids (pSB3K3) using proteogenic amino acids and milk as organocatalysts DNA vectors for multi-step, combinatorial gene repression/knockout (KO) recipes for DNA origami forensic detection (N-P bound phosphoramidate), and exogenous drugs or antiviral proteins expressions with high bioluminiscence activity:
- Orthogonal phosphorus-degrading enzymes are key in (auto)catalytic regulation of lacZ-bound phosphate-histidine residues - see N-methyl nucleosidase (220.127.116.11) and this page for references.
In this experiment the goal is to construct a forensic DNA fingerprinting kit to verify the DNA fragments for evidences of electrocompetent-inducible promoters and cross-species gene vectors to produce bioluminiscence agaisnt a control group (MG1655).
- Control group will need to be plasmid-free - Obtain E. coli strain mg1655 as reference model for the electrophoresis experiment.
- DNA extraction: data interpretation is subject to errors in the protocol - Find a coherent protocol specific for E.coli transformations using multi-plasmid vectors to transform E.coli into the desired mutant strain.
- familiarize yourself with the various electrophoresis assay methods - An important resource to learn and study the basics of E.coli transcription ab initio using recombinant DNA.
- more checklist fine points here...
- E. coli restriction-modification system
- Standard E. coli Strain for BioBricks - and this paper
- Start up genome engineering [with the M13 phage genome] - [a picture is HERE]
- Beta-galactosidase assay - EC 18.104.22.168
- sequence for M13k07 reengineered plasmid : Note : If pMR101 is a gene vector derived from pACYC177 then clearly the resulting plasmid cloning vector will be a synthetic construct derived from a M13 like bacteriophage, thus the use of restriction enzymes for acquiring inter-species genes (OmpR/EnvZ) is likely a positive indicator of a scheme to induce horizontal gene regulation using bacterial conjugation like sex in elephants and the E. coli K-12 MG1655 strain.
- The Matrix (1999)