Difference between revisions of "Part:BBa K1781000"

Revision as of 22:12, 16 September 2015

P3 from M13

M13 gene 3 coding for infection protein P3

Sequence and Features

Functional Parameters

Some difficulties were found with the expression of P3 and the resulting washout of the phages. The plasmid showed itself stable during the whole cultivation which makes the next step a cultivation without antibiotics possible and highly desirable.

Further experiments regarding the plasmid stability should be performed, as well as reducing the yeast extract from the medium by identifying the missing element in the minimal medium. A special expression analysis has to be done on P3 in relation to the IPTG-concentration.

In order to ensure proper antibiotic denaturation the cultivated medium had to be sterilized for 20 minutes at 121 °C. Further experiments are necessary to test the need of antibiotics for plasmid stability in the system. Since the antibiotic resistance of the recombinant E. coli strand is based on chloramphenicol degradation..

Additionally, the synthesized enzyme is secreted into the cultivation medium. This might lead to a complete loss of antibiotic function and therefore allow plasmid free E. coli to reproduce. As a result, the plasmid free cells might accumulate inside the CSR. Thus it is necessary to compare the plasmid stability of antibiotic free and antibiotic enriched media. Accounting to the previous reasons the influence of the chloramphenicol on plasmid stability might be negligible. As a result other ways to support plasmid stability or antibiotic free systems might be used for further experiments.