Team:Michigan/Results

From 2012.igem.org

(Difference between revisions)
Line 30: Line 30:
<br>
<br>
-
<br>
+
<br><br>
</html>
</html>

Revision as of 00:15, 4 October 2012

Xayona Website Template


Introduction

The Asymmetrical Digest Reporters

In order to test the effectiveness of the recombinases used in our system, we developed two assays -- an “asymmetrical digest” reporter and a fluorescence reporter. The former is used to directly test the inversion of the target DNA region, while the latter is used to demonstrate inversion induced protein expression.

The Fluorescent Reporters

The two asymmetrical digest reporters in their unflipped state (invertible by FimE) are shown in figures 2 and 3. These reporters rely on an off-center AgeI restriction site which will yield fragments of varying size depending on the orientation of the inversion region. Table 1 shows the expected sizes of the fragments in each orientation. The differences between the reporters are discussed below.

Figure 1. A diagram of the asymmetrical reporter K880001




Figure 2. A diagram of the asymmetrical reporter K880003

Results

Negative Fluorescent Reporter (BBa_K137058) Data

Negative Asymmetrically Digestible Reporter (BBa_K880003) Data

Observation of IRR and IRL Orientation:

Modified Asymmetrically Digestible Reporter (BBa_K880001) Data:

Negative Modified Fluorescent Reporter (BBa_K880002) Data:

Observation of HbiF induced fimbriation in DH5alpha