Part:BBa_K1996002: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_K1996002

EtnR1 protein from Mycobacterium NBB4, when expressed in E. coli. and purified by his-tag, binds to the EtnP promoter (<a href="https://parts.igem.org/Part:BBa_K1996000">BBa_K1996000</a>) as demonstrated by electrophoretic mobility shift assay (Figure 1). When bound by EtnR1, the EtnP DNA is less able to migrate through the agarose gel and therefore presents at a higher band size.

<img src="" alt="EtnR1 EMSA" width="400" height=:300">
Figure 1. Electrophoretic mobility shift assay for EtnP DNA. The 250 bp region of EtnP was PCR amplified and 100 ng of DNA incubated with 6 mg of protein. The protein-DNA mixtures were run on a 3% agarose gel at 150 V for 100 minutes and stained with Gel Green overnight for visualisation. Lane 1: NEB 100 bp ladder. Lane 2: EtnP DNA only. Lane 3: EtnR1 protein only. Lane 4: EtnR1 protein and EtnP DNA. Lane 5: BSA protein and EtnP DNA.

User Reviews

UNIQ1fd895ffd0fcd485-partinfo-00000000-QINU UNIQ1fd895ffd0fcd485-partinfo-00000001-QINU