Part:BBa_K419003:Experience

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_K419003

Verification of the Part BBa_k419003

The DNA fragment of LuxR responsive promoter, LuxPR, was amplified from the part BBa_R0062 by PCR method. The LuxPR PCR products were purified and digested with EcoRI and PstI, then ligated with EcoRI-PstI cut pSB1C3, the iGEM 2010 standard backbone. We got the pSB1C3-LuxPR plasmid successfully. In addition, the lysis device DNA which contained holing and lysozyme genes was amplified from the part BBa_K112021. The lysis device PCR products were digested with Xba I and Pst I and ligated with the Spe I-Pst I treated pSB1C3-LuxPR vector. After transformation and plasmid preparation, the part BBa_k419003 for releasing system in our Neutrient Synthesizer was verified by EcoRI and PstI enzyme digestion in 1% agarose gel electrophoresis. The arrow (red) indicates the lysis device DNA fragment. The releasing system composite was transformed into DH5-Alpha E. coli for further experiment.

Demonstration of Releasing System [contains the part BBa_k419003 (pLuxPR-Lysis device) in the iGEM 2010 standard backbone pSB1C3]

A single colony of releasing system E. coli was inoculated in the LB/chloramphenicol medium at 37℃. After 8 h of culture, we found that the LuxPR was auto-activated and lysis device was expressed causing the lysis of E. coli host. In our observation, we found that the autolysis of the host bacterial was almost completed after 8 hours. The result indicated that other factors may be involved in the activation of the luxR responsive promoter.

User Reviews

UNIQ09554d153885d948-partinfo-00000000-QINU UNIQ09554d153885d948-partinfo-00000001-QINU