Team:Heidelberg/Templates/Indigoidine week6 overview

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===Indigoidine Production - bpsA===
===Indigoidine Production - bpsA===
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This week we repeated the co-transformation of both the [[Strain#Rosetta|''E. coli'' Rosetta]] and the BAP1 strain with the bpsA ([[Plasmids#pMM64|pMM64]]) and the svp ([[Plasmids#pMM65|pMM65]]) plasmids. The [[Strain#BAP1|''E. coli'' BAP1]] strain is a variant of [[Strain#BL21DE3|''E. coli'' BL21(DE3)]] and contains the sfp PPTase from [[Strain#Bsub168|''B. subtilis'' 168]] (<bib id="Nakano1992"/><bib id ="Quadri1998"/>) and a T7-promoter, brought in using a genomic integration approach (<bib id ="Pfeifer2001"/>). The Pfeifer lab kindly supported us by sending the BAP1 strain.<br/>
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This week we repeated the co-transformation of both the ''E. coli'' Rosetta and the BAP1 strain with the bpsA (pMM64) and the svp (pMM65) plasmids. The ''E. coli'' BAP1 strain is a variant of ''E. coli'' BL21(DE3) and contains the sfp PPTase from ''B. subtilis'' 168 (Nakano 1992, Quadri 1998) and a T7-promoter, brought in using a genomic integration approach (Pfeifer 2001). The Pfeifer lab kindly supported us by sending the BAP1 strain.<br/>
After induction, liquid cultures of positive BAP1 transformants should turn blue.
After induction, liquid cultures of positive BAP1 transformants should turn blue.
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Latest revision as of 15:06, 21 October 2013

Indigoidine Production - bpsA

This week we repeated the co-transformation of both the E. coli Rosetta and the BAP1 strain with the bpsA (pMM64) and the svp (pMM65) plasmids. The E. coli BAP1 strain is a variant of E. coli BL21(DE3) and contains the sfp PPTase from B. subtilis 168 (Nakano 1992, Quadri 1998) and a T7-promoter, brought in using a genomic integration approach (Pfeifer 2001). The Pfeifer lab kindly supported us by sending the BAP1 strain.
After induction, liquid cultures of positive BAP1 transformants should turn blue.

Both strains still didn't turn blue when grown and induced on plate and in liquid culture, even after some days with different growth conditions. We continued with the validation of the pMM-plasmids.