Difference between revisions of "Part:BBa K3733005"
Heping Zhang (Talk | contribs) |
|||
| Line 3: | Line 3: | ||
<partinfo>BBa_K3733005 short</partinfo> | <partinfo>BBa_K3733005 short</partinfo> | ||
| − | + | <p>J23106a is a constitutive promoter obtained by mutating <a url="https://parts.igem.org/Part:BBa_J23106"> J23106</a>. The strength of this promoter is much weaker than J23106. </p> | |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | <p>J23106 is a medium-strength constitutive promoter of the Anderson family. By editing J23106's -35 region by replacing nucleotides 3-5 from TAC to ATA, J23106a was successfully constructed (<b>Figure 1</b>)，which is a very weak promoter. </p> | |
| + | <div><img></img></div> | ||
| + | <p><b>Figure 1. </b> Modified methods of J23106a</p> | ||
<!-- --> | <!-- --> | ||
| + | |||
| + | ===Functional Parameters=== | ||
| + | <p>To characterize this part, J23106a and J23106 were cloned into pSC101 vector separately. We chose <a url="https://parts.igem.org/Part:BBa_K3733012">neGFP </a> as the reporter. Plasmids were transferred into <i>E.coli</i> DH5α. The E. coli strain was cultured overnight, and the relative intensity of J23106a was evaluated by measuring RFU/OD<sub>600</sub> under the control of two different promoters. The relative fluorescence unit (RFU) was determined by Synergy H1 microplate reader. </p> | ||
| + | <div><img></img></div> | ||
| + | <p><b>Figure 2. </b> Comparison of relative strength between J23106 and J23106a.The microplate reader gain adopts 50</p> | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K3733005 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3733005 SequenceAndFeatures</partinfo> | ||
Revision as of 00:48, 15 October 2021
J23106a:Improved promoter of J23106
J23106a is a constitutive promoter obtained by mutating <a url="https://parts.igem.org/Part:BBa_J23106"> J23106</a>. The strength of this promoter is much weaker than J23106.
Functional Parameters
To characterize this part, J23106a and J23106 were cloned into pSC101 vector separately. We chose <a url="https://parts.igem.org/Part:BBa_K3733012">neGFP </a> as the reporter. Plasmids were transferred into E.coli DH5α. The E. coli strain was cultured overnight, and the relative intensity of J23106a was evaluated by measuring RFU/OD600 under the control of two different promoters. The relative fluorescence unit (RFU) was determined by Synergy H1 microplate reader.
Figure 2. Comparison of relative strength between J23106 and J23106a.The microplate reader gain adopts 50
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]