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During our project we tried to address issues that arose during the Washington 2009's project. After preforming sequence analysis, it seemed that for DH5Alpha cells the native ribosomal binding sites may not be as easily recognized by the ribosomes. This would cause decreased cellular expression. Similarly analysis suggested that the parts would have benefited from codon optimization transferring the protein from a plant pathogen to E.coli. We are currently working on optimizing the sequence for both DH5alpha and E.coli Nissle 1917 our chassis organism. Furthermore, according to the dual plasmid system a dual selector would be required, this would cause added stress to the cell diminishing the synthesis of the pump to be inserted into the membrane as well as diminish the synthesis of the tagged proetin of interest to be exported out of the cell. This could be counteracted with a super optimum media.

Revision as of 03:06, 28 September 2013

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_K215104

This BioBrick contains the three essential parts of the Erwinia chrysanthemi Type I secretion system.

User Reviews

This part was used to make the following BioBrick: BBa_K215107.

UNIQ598a2b2f862e1314-partinfo-00000000-QINU UNIQ598a2b2f862e1314-partinfo-00000001-QINU

During our project we tried to address issues that arose during the Washington 2009's project. After preforming sequence analysis, it seemed that for DH5Alpha cells the native ribosomal binding sites may not be as easily recognized by the ribosomes. This would cause decreased cellular expression. Similarly analysis suggested that the parts would have benefited from codon optimization transferring the protein from a plant pathogen to E.coli. We are currently working on optimizing the sequence for both DH5alpha and E.coli Nissle 1917 our chassis organism. Furthermore, according to the dual plasmid system a dual selector would be required, this would cause added stress to the cell diminishing the synthesis of the pump to be inserted into the membrane as well as diminish the synthesis of the tagged proetin of interest to be exported out of the cell. This could be counteracted with a super optimum media.