Part:BBa_K3745030:Design
Plac+rhlABC linear fragment
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 3524
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2185
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2149
Illegal NgoMIV site found at 2458
Illegal NgoMIV site found at 4067
Illegal AgeI site found at 3070 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 4206
Design Notes
It is worth informing that the Plac in the DNA feature's graph is a reverse promoter (activate LacI instead of activating rhlA, B, and C). The arrow on the feature is not accurate due to the reason that the image of the promoter won't appear in the reverse direction. Nevertheless, the DNA sequence is posted in the correct arrangement which Plac is responsible for activating LacI.
Source
This study
Producing and Quantifying Rhamnolipid
Our team aims to produce rhamnolipid by using BBa_K2745030 plasmid including Plac, rhlA, rhlB, and rhlC. First, IPTG is involved in our production process due to the reason that IPTG is a substance that is similar to lactose. In other words, it can be used to induce Plac and futher propel the production of rhamnolipids by measure the amount of production of rhamnolipids by using the HPLC-MS method. The specific quantification steps are shown below. Quantification using HPLC-MS 1. Add HCl into the liquid culture until the final pH is 2 2. Leave the solution overnight at 4° C 3. Extract the organic phase using an ethanol-chloroform solution in a ratio of 1:2. 4. HPLC-MS is performed using C18 column and acetonitrile as mobile phase.