Difference between revisions of "Part:BBa K2268004"

Latest revision as of 02:19, 2 November 2017

p(cI)-LacZ alpha, [p(Lac)-cI] reverse

This is the LacZ alpha fragment (BBa_I732006) under the control of a cI responsive promoter (BBa_R0051), followed by a reverse complement copy of the cI repressor (BBa_K2268000) under the control of a LacI responsive promoter (BBa_K2268001). The latter two components act as an inverter: addition of IPTG will repress expression of the LacZ reporter, while a lack of IPTG will lead to expression.

LacZ alpha fragment can associate with the omega fragment, if present, to restore beta-galactosidase activity to a cell. This can be visualized as a blue pigment color when cells are grown in the presence of X-gal.

Note: the Sequence diagram below does not reflect the promoter, BBa_K2268001, that drives expression of the cI repressor (K2268000) for technical reasons.

Usage and Biology

When grown in the presence of X-gal, the LacZ alpha fragment gene (regulated at the promoter by the cI repressor) will lead to a blue pigment to accumulate in the bacteria. Here, shown below are images of K2268003 and K2268004, co-transformed into E. coli with pDawn, which produces the cI repressor under dark conditions (which thereby shuts down LacZ expression). Images are shown to indicate expression in the absence or presence of IPTG (relevant for induction of supplementary cI repressor in construct K2268004).

Sequence and Features

More information on the construction of this part, and the sequencing results, can be found at the Kingsborough 2017 iGEM team Wiki (Relevant page: http://2017.igem.org/Team:Kingsborough_NY/BioBrick)