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Team:LCG-UNAM-Mexico:Journals:Uriel
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(New page: == 31 Jul 2009 == To attain control of phage production we are going to put under an IPTG inducible promoter the genes that codify for the principal regulators of the morphopoietic gen...)
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31 Jul 2009
To attain control of phage production we are going to put under an IPTG inducible promoter the genes that codify for the principal regulators of the morphopoietic genes.
The genes that codify for this regulators are going to be amplified via colony PCR from the E. coli C-177 strain, which has a lysogenic form of phage P2. The primers that e are using amplify the genes with its WT RBS's and also introduce an extra stop codon as required by the standardization protocol and also de suffix and prefix iGEM sequences.
It is known that C-1a strain neither has a cox gene nor ogr gene while K-12 strain contain a copy of ogr in its genome. We have performed a colony PCR over the next strains, looking for ogr and cox.
C-1a C-117 DH5alpha
PCR Reaction:
PCR:
Control - C-117/ogr + C-1a/ogr - DH5alpha/ogr + C-117/cox + C-1a/cox - DH5alpha/cox -
The PCR's were consistent with previous results in literature. For example ogr was positive for DH5alpha which is a derivative from K-12 and we could not obtain PCR products from C-1a. C-117 which has P2 in lysogenic state gave use positive PCR products for ogr and cox.
