Difference between revisions of "Part:BBa K942004:Experience"

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===Applications of BBa_K942004===
 
===Applications of BBa_K942004===
  
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This protein was expressed in Pichia pastoris. Thanks to its B.P.S. it was also planned to be expressed in Escherichia coli. We succesfully transformed the gene into our cloning strains, but curiously, our expression strains (DE3 strains) were not even able to be transformed, except from Rosettagami DE3 pLys. We believe that the fact that Rosettagami had pLys to prevent leak expression and the other expression strains did not, leads us to the idea that there might be posibility of Der f 2 having antibacterial effects, as it is describe on the main page of this same part.
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Our shuttle sequence can be quite useful to other iGEM teams and anyone with the objective of producing or comparing the expression of proteins in two different expression systems. Coupled with an codon optimization for Pichia p. and a strain capable of reading rare codons like Rosetta Gami, Bl21 star, the outcome can be a simultanious expression in both organisms coming from one single genetic sequence
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We were able to se the expression of this protein on a SDS-PAGE after purifying it with a His-tag affinity column.
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Currently, we are trying different conditions to achieve the expression in BL21 Star.
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===User Reviews===
 
===User Reviews===

Revision as of 21:00, 28 September 2012

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K942004

Our shuttle sequence can be quite useful to other iGEM teams and anyone with the objective of producing or comparing the expression of proteins in two different expression systems. Coupled with an codon optimization for Pichia p. and a strain capable of reading rare codons like Rosetta Gami, Bl21 star, the outcome can be a simultanious expression in both organisms coming from one single genetic sequence

User Reviews

UNIQ353a05a959b8d525-partinfo-00000000-QINU UNIQ353a05a959b8d525-partinfo-00000001-QINU


This protein was expressed in Pichia pastoris. Thanks to its B.P.S. it was also planned to be expressed in Escherichia coli. We succesfully transformed the gene into our cloning strains, but curiously, our expression strains (DE3 strains) were not even able to be transformed, except from Rosettagami DE3 pLys. We believe that the fact that Rosettagami had pLys to prevent leak expression and the other expression strains did not, leads us to the idea that there might be posibility of Der f 2 having antibacterial effects, as it is describe on the main page of this same part. We were able to se the expression of this protein on a SDS-PAGE after purifying it with a His-tag affinity column. Currently, we are trying different conditions to achieve the expression in BL21 Star.