Revision as of 07:27, 12 November 2012

Plasmid cloning vectors for gene therapy with phosphohistidine-based oligos

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:

Orthogonal DNA shell with Benzyl-derived (Bz) protecting groups:

note! note! Important: Can you elucidate the reasoning of using an carboxilic acid as protecting group for DNA assembly?

References:

@@ Line 23: / Line 23: @@
 [http://openwetware.org/wiki/20.109(S07):Start-up_genome_engineering#Part_3:_Digest_M13K07 M13K07] cloning in ''E. coli'':
 * amino-acid sequence signature (Myc epitope tag) = EQKLISEEDL
+Orthogonal DNA shell with Benzyl-derived (Bz) [http://en.wikipedia.org/wiki/Protecting_group#Carboxylic_acid_protecting_groups protecting groups]:
+* note! note! Important: Can you elucidate the reasoning of using an carboxilic acid as protecting group for DNA assembly?
 References: