Difference between revisions of "Part:BBa K1996001"
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Putative transcription factor for enzymes involved in ethylene degradation pathway in Mycobacterium NBB4 | Putative transcription factor for enzymes involved in ethylene degradation pathway in Mycobacterium NBB4 | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| + | EtnR1 is a DNA-binding protein that binds to the EtnP (BBa_K1996000) sequence. It is believed that this regulates the expression of ethylene-oxidising genes. In an electrophoretic mobility shift assay (EMSA), it was found that EtnR1 binding to EtnP notably inhibits the migration of the DNA through the gel, indicating a strong binding affinity between EtnR1 protein and EtnP DNA (Figure 1). | ||
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| + | <img src="https://static.igem.org/mediawiki/2016/b/b0/T--Sydney_Australia--GelShiftFigure.png"><br> | ||
| + | <b>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. </b> | ||
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Revision as of 01:42, 21 October 2016
EtnR1 - DNA-binding protein for EtnP in Mycobacterium NBB4
Putative transcription factor for enzymes involved in ethylene degradation pathway in Mycobacterium NBB4
Usage and Biology
EtnR1 is a DNA-binding protein that binds to the EtnP (BBa_K1996000) sequence. It is believed that this regulates the expression of ethylene-oxidising genes. In an electrophoretic mobility shift assay (EMSA), it was found that EtnR1 binding to EtnP notably inhibits the migration of the DNA through the gel, indicating a strong binding affinity between EtnR1 protein and EtnP DNA (Figure 1).
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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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 150
Illegal AgeI site found at 1359 - 1000COMPATIBLE WITH RFC[1000]