User:Etienne Robillard/Notebook/Agent Ecoli:Plasmid cloning vectors for gene therapy with phosphohistidine based oligos

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(Artificial DNA primers: phosphoramidate-based oligonucleotide synthesis)
(Artificial DNA primers: phosphoramidate-based oligonucleotide synthesis)
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** P = a Phosphonate (inorganic state) - a compound for oligo synthesis using the [http://en.wikipedia.org/wiki/Oligonucleotide_synthesis#Nucleoside_phosphoramidites nucleoside phosphoramidite] method.   
** P = a Phosphonate (inorganic state) - a compound for oligo synthesis using the [http://en.wikipedia.org/wiki/Oligonucleotide_synthesis#Nucleoside_phosphoramidites nucleoside phosphoramidite] method.   
** P-N = phosphoramidate (a Nitrogen atom bound to a Phosphonate ester in nucleophilic substitution).
** P-N = phosphoramidate (a Nitrogen atom bound to a Phosphonate ester in nucleophilic substitution).
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deoxynucleotide synthesis:
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* cDNA synthesis building block = deoxynucleoside + 3'-phosphonate = synthetic nucleotide analog (dATP) [http://dwb4.unl.edu/Teacher/NSF/C08/C08Links/www.dur.ac.uk/~dbl0www/Staff/Croy/cDNAfigs.htm 1]
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oligodeoxyribonucleotide synthesis:
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* short 50-200(?) bp restriction endonucleases enzymes for cDNA catalysis/plasmid vector cloning in ''E. coli''
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* building blocks for oligonucleotides synthesis with phosphoramidites method [http://en.wikipedia.org/wiki/Oligonucleotide 1]
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* see also [http://openwetware.org/wiki/CH391L/S12/GeneandGenomeSynthesis#Oligonucleotide_Synthesis Oligonucleotide Synthesis]
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[http://openwetware.org/wiki/20.109(S07):Start-up_genome_engineering#Part_3:_Digest_M13K07 M13K07] cloning in ''E. coli'':
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* amino-acid sequence signature (Myc epitope tag) = EQKLISEEDL
== References ==
== References ==

Revision as of 04:23, 9 November 2012

Plasmid cloning vectors for gene therapy with phosphohistidine-based oligos

Contents

Artificial DNA primers: 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 synthesis with phosphoramidites method 1
  • see also Oligonucleotide Synthesis

M13K07 cloning in E. coli:

  • amino-acid sequence signature (Myc epitope tag) = EQKLISEEDL

References

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