Difference between revisions of "Part:BBa J179000"

Latest revision as of 16:55, 3 February 2014

This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the amyE locus and can be selected with Chloramphenicol (cat gene). It has a ampicillin resistance for cloning in E.coli. This backbone is a BioBricked version of the B. subtilis vector pDG1662 ([http://www.ncbi.nlm.nih.gov/pubmed/8973347 Guérout-Fleury et al.]).

This version is derived from BBa_K823023 and does not contain AgeI nor NgoMIV restriction sites, therefore is RFC25-compatible.

This part was also evaluated in the publication [http://www.jbioleng.org/content/7/1/29 The Bacillus BioBrick Box: generation and evaluation of essential genetic building blocks for standardized work with Bacillus subtilis] by Radeck et al..

This BioBrick is part of the [http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks BacillusBioBrickBox] developed by the LMU-Munich 2012 iGem Team.

For handling of B. subtilis vectors, please see here.

The transformation into B. subtilis is explained here.

To check for integration of this vector into the amyE locus, the starch test can be used. The colonies obtained from the transformation into B.subtilis were streaked on an LB-Agar plate with 5 mg/L chloramphenicol and onto a starch plate (also W168 as a control) and incubated at 37°C over night. The starch plate is then covered with Lugol's iodine. Colonies without a bright surrounding are correct. We tested this vector with various inserts, for example the final construct of our Sporobeads with GFP. The starch test shows that the insertion in in the amyE locus and fluorescence microscopy reveals the functionality of the construct.

Sequence and Features