User:Etienne Robillard/Notebook/Agent Ecoli:SolidPhasePeptideSynthesisReagentEvidencesInAgentScoobyDoo
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(New page: == Measuring the amplitude of the synthetic reagent lysis TE buffer on NMDA receptor == * Date: 2012/12/03 10:29 AM EDT * Organism: E. coli * Strain: === Hypothesis === Amine protec...) |
Current revision (21:01, 12 December 2012) (view source) (→Hypothesis) |
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* Date: 2012/12/03 10:29 AM EDT | * Date: 2012/12/03 10:29 AM EDT | ||
| - | * Organism: E. coli | + | * Organism: E. coli (K-12 substr.) |
| - | * Strain: | + | * Strain: [http://cgsc2.biology.yale.edu/Strain.php?ID=65553 BW26434] |
| + | |||
=== Hypothesis === | === Hypothesis === | ||
| - | Amine protection groups (piperidine based) undergo chiral nucleophilic substitution on Histidine based autophosphorylation. | + | * Amine protection groups (piperidine based) undergo chiral nucleophilic substitution on Histidine based autophosphorylation. |
| + | * Cosmid like recombinase system for -35 lacl+ (LacZ) transcription, thus no infection produced; (no lacY) | ||
| + | * orthogonal oligo/peptide protection using methylamino (methyltransferase) agents; | ||
| + | * methylamino buffering agents used as intermediates for orthogonal protection; (probably a side-effect of Magnesium chelation with EDTA?) | ||
| + | * artificial cos-mid size is 30-45 bp length => reduced genome ? | ||
| + | * The NMDA receptor is voltage gated: [http://www.physicsforums.com/showthread.php?t=632781 The addition of EDTA (mg2+) ions to the reaction may act as '''NMDA receptor antagonist''' by binding to the NMDA ligand...] | ||
| + | * [[IPTG]] induction is a chemical reaction which may be neutralized with 1mL NaHC03... (Sodium bicarbonate, 2.99$) [[http://en.wikipedia.org/wiki/Sodium_bicarbonate#Neutralization_of_acids_and_bases 1]] [[http://bioserv.fiu.edu/~biolab/labs/genetics/lac%20operon.htm 2]] [[http://www.openwetware.org/wiki/BE.109:Protein_engineering/Assessing_beta-galactosidase 3]] [[http://www.openwetware.org/wiki/Beta-Galactosidase_Assay_(A_better_Miller) 4]] | ||
=== Experiment === | === Experiment === | ||
| Line 18: | Line 25: | ||
* '''Amine''' protection groups = piperidine derived compound (NH2 undergo nucleophilic substitution on Histidine based autophosphorylation) | * '''Amine''' protection groups = piperidine derived compound (NH2 undergo nucleophilic substitution on Histidine based autophosphorylation) | ||
| - | + | * a heterocyclic base-labile group (NHC?) | |
| + | * [[IPTG]] inducible promoter: DNA lambda prophage induction (Transformation or mutation from lytic lacZ-repressed state to lysogenic!) | ||
| + | ** in that case a matching E. coli genotype is "HB101 lacIq, lacY+" (ie [http://partsregistry.org/Part:BBa_V1003 laclq]) | ||
| + | ** From [http://www.sigmaaldrich.com/life-science/molecular-biology/cloning-and-expression/learning-center/genotypes-of-e-coli.html Sigma], the expected genotype matching the laclq strain is: | ||
| + | *** HB101 F– Δ(gpt-proA)62 leuB6 glnV44 ara-14 galK2 lacY1 Δ(mcrC-mrr) rpsL20 (Strr) xyl-5 mtl-1 recA13h | ||
| + | * (2R)-2-(methylamino)succinic acid - An orthogonal N-methyltransferase/NH2 base labile compound; exogenous NMDA receptor antagonist compound (Ketamine-like); synthetically recombined oligonucleotide protection group (dA-dT) for solid-phase peptide synthesis (SPPS), etc (see also [[PHP]], [[Succinic_acid]], and BW26434.) | ||
| + | ** important: N-methyl-D-asparte receptor is a "sensory" neurotransmitter with analogous functions to the TRPV neurotransmitter but for brain functions including learning and memory. | ||
==== References ==== | ==== References ==== | ||
| - | * [TE] | + | * [https://www.openwetware.org/wiki/TE TE] |
| - | * [ | + | * [http://cgsc.biology.yale.edu/Strain.php?ID=9130 AB266] |
| + | * [http://cgsc.biology.yale.edu/Strain.php?ID=11958 E.coli B]: WOOP WOOP | ||
| + | * [[IPTG]] | ||
| + | * [[EDTA]] | ||
Current revision
Contents |
Measuring the amplitude of the synthetic reagent lysis TE buffer on NMDA receptor
- Date: 2012/12/03 10:29 AM EDT
- Organism: E. coli (K-12 substr.)
- Strain: BW26434
Hypothesis
- Amine protection groups (piperidine based) undergo chiral nucleophilic substitution on Histidine based autophosphorylation.
- Cosmid like recombinase system for -35 lacl+ (LacZ) transcription, thus no infection produced; (no lacY)
- orthogonal oligo/peptide protection using methylamino (methyltransferase) agents;
- methylamino buffering agents used as intermediates for orthogonal protection; (probably a side-effect of Magnesium chelation with EDTA?)
- artificial cos-mid size is 30-45 bp length => reduced genome ?
- The NMDA receptor is voltage gated: The addition of EDTA (mg2+) ions to the reaction may act as NMDA receptor antagonist by binding to the NMDA ligand...
- IPTG induction is a chemical reaction which may be neutralized with 1mL NaHC03... (Sodium bicarbonate, 2.99$) [1] [2] [3] [4]
Experiment
Material
- 1 x 10ml TE = TAE -> 10x buffer (Tris-HCl) (see [TE]
Expected results
- Amine protection groups = piperidine derived compound (NH2 undergo nucleophilic substitution on Histidine based autophosphorylation)
- a heterocyclic base-labile group (NHC?)
- IPTG inducible promoter: DNA lambda prophage induction (Transformation or mutation from lytic lacZ-repressed state to lysogenic!)
- (2R)-2-(methylamino)succinic acid - An orthogonal N-methyltransferase/NH2 base labile compound; exogenous NMDA receptor antagonist compound (Ketamine-like); synthetically recombined oligonucleotide protection group (dA-dT) for solid-phase peptide synthesis (SPPS), etc (see also PHP, Succinic_acid, and BW26434.)
- important: N-methyl-D-asparte receptor is a "sensory" neurotransmitter with analogous functions to the TRPV neurotransmitter but for brain functions including learning and memory.
