Team:NU Kazakhstan/Parts
Background
Overview
Our team added rhlA/rhlB and nadE genes to the pRGPDuo2 plasmid. The pRGPDuo2 plasmid was obtained from Gauttam, R. We extracted rhlA/rhlB and nadE genes from P.aeruginosa. Our team planned to insert rhlA and rhlB genes to MCS1 and nadE gene to MCS2 of pRGPDuo2 plasmid. Thus, we predicted that dual expression of NAD synthetase and RhlA and RhlB could allow P. putida to express rhamnolipids at higher rates. Overall, we created ten basic parts (four coding sequences and six primers) and one composite part.
Parts List
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K4083004 | Coding | nadE gene of Pseudomonas aeruginosa | Arsen Orazbek | 883 bp |
| BBa_K4083006 | Coding | rhlB with SacI and SalI sites | Arsen Orazbek | 1334 bp |
| BBa_K4083007 | Coding | rhlA gene with SalI and SacI sites at ends | Arsen Orazbek | 932 bp |
| BBa_K4083000 | Plasmid | pRGPDuo2 plasmid for P. putida | Arsen Orazbek | 7928 bp |
| BBa_K4083008 | Composite | RPGDuo2 plasmid with nadE and rhlA/B genes | Arsen Orazbek | 6403 bp |
We used the following primers:
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K4083020 | Primer | Reverse primer for nadE | Arsen Orazbek | 30 bp |
| BBa_K4083019 | Primer | Forward primer for nadE | Arsen Orazbek | 31 bp |
| BBa_K4083018 | Primer | Reverse primer for rhlB | Arsen Orazbek | 37 bp |
| BBa_K4083016 | Primer | Forward primer for |
Arsen Orazbek | 38 bp |
| BBa_K4083015 | Primer | Primer for rhlA reverse | Arsen Orazbek | 37 bp |
| BBa_K4083014 | Primer | Forward Primer for rhlA | Arsen Orazbek | 36 bp |
Although we obtained pRGPduo2 plasmid already constructed, we created and used several parts for BioBricks construction purposes.
Reference List:
Gauttam, R., Mukhopadhyay, A., & Singer, S. W. (2020). Construction of a novel dual-inducible duet-expression system for gene (over)expression in Pseudomonas putida. Plasmid, 110. https://doi.org/10.1016/j.plasmid.2020.102514
CONTACT
igem@nu.edu.kz
Kabanbay batyr av., 53, Nur-Sultan, Kazakhstan
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