User:Etienne Robillard/Notebook/Chemtrails911 notebook/Agent Ecoli:Plasmid cloning vectors for gene therapy with phosphohistidine based oligos
From OpenWetWare
(Difference between revisions)
(→Artificial DNA primers: phosphoramidate-based oligonucleotide synthesis) |
(→Artificial DNA promoters: phosphoramidate-based oligonucleotide synthesis) |
||
| Line 19: | Line 19: | ||
* building blocks for oligonucleotides assembly with phosphoramidites method [http://en.wikipedia.org/wiki/Oligonucleotide 1] | * building blocks for oligonucleotides assembly with phosphoramidites method [http://en.wikipedia.org/wiki/Oligonucleotide 1] | ||
* see also [http://openwetware.org/wiki/CH391L/S12/GeneandGenomeSynthesis#Oligonucleotide_Synthesis Oligonucleotide Synthesis] | * see also [http://openwetware.org/wiki/CH391L/S12/GeneandGenomeSynthesis#Oligonucleotide_Synthesis Oligonucleotide Synthesis] | ||
| + | * 1 mutation, 1 oligo assay (phosphoramidite-based oligo synthesis): [http://openwetware.org/wiki/Reviews/Directed_evolution/Library_construction#Directing_diversity:_Oligonucleotide-based_methods] | ||
[http://openwetware.org/wiki/20.109(S07):Start-up_genome_engineering#Part_3:_Digest_M13K07 M13K07] cloning in ''E. coli'': | [http://openwetware.org/wiki/20.109(S07):Start-up_genome_engineering#Part_3:_Digest_M13K07 M13K07] cloning in ''E. coli'': | ||
Revision as of 06:52, 12 November 2012
Plasmid cloning vectors for gene therapy with phosphohistidine-based oligos
Contents |
Artificial DNA promoters: phosphoramidate-based oligonucleotide synthesis
cDNA:
- N-6-benzoyl-deoxyadenosine phosphoramidite - Notice the P-N bound indicating the linkage of a phosphonate diester to a benzoyl atom :)
- N-3-oxo-hexanoyl-L-homoserine lactone - A light-inducible phosphohistidine-bound AHL ? A(denosine)TP/cAMP dependent.
- Reminder:
- NHC = (N heterocycle carbene) - Chemical compounds with a N chiral prodrug switch - highly enantioselective molecules
- N = indicates a Nitrogen atom
- P = a Phosphonate (inorganic state) - a compound for oligo synthesis using the nucleoside phosphoramidite method.
- P-N = phosphoramidate (a Nitrogen atom bound to a Phosphonate ester in nucleophilic substitution).
deoxynucleotide synthesis:
- cDNA synthesis building block = deoxynucleoside + 3'-phosphonate = synthetic nucleotide analog (dATP) 1
oligodeoxyribonucleotide synthesis:
- short 50-200(?) bp restriction endonucleases enzymes for cDNA catalysis/plasmid vector cloning in E. coli
- building blocks for oligonucleotides assembly with phosphoramidites method 1
- see also Oligonucleotide Synthesis
- 1 mutation, 1 oligo assay (phosphoramidite-based oligo synthesis): [1]
M13K07 cloning in E. coli:
- amino-acid sequence signature (Myc epitope tag) = EQKLISEEDL
References:
