Part:BBa_K3895012
Insecticidal protein
Bt toxins refer to the toxic proteins produced by insect pathogenic bacteria Bacillus thuringiensis[1]. This part was adjusted from BBa_K3686010 (BBa_K3686010), where Cry7Ca1 is a differentiate of Bt toxins with a molecular mass of 129kDa, recently isolated from Bt strain BHT-13. Cry7Ca1 is connected with GFP (BBa_E0040),and constructed into pCAMBIA-Cry7Ca1-eGFP vector.
Protocol
1. Transfection of pCAMBIA-Cry7Ca1-eGFP vector through Carbon dots nanocomposite (CDP) to Nicotiana benthamiana
Coupling CDP with plant expressing vector Mix the following ingredients:
| Ingredients | Volumn |
|---|---|
| MES buffer(50X) | 10ul |
| CDP(50X) | 10ul |
| DNA(237ng/ul >10ng/ul final con) | 22ul |
| 10% glycerol | 25ul |
| ddH2O | 433ul |
| total | 500ul |
Gently mix and incubate at 37℃ for 30min.
2. Brush the mixture gently on the leaf of Nicotiana benthamiana(leaf length>10cm,Growing well) ，mark the area of brushing
3. Put the plant back in the light incubator(28℃，12h light 12h dark), repeat the process for four days at 3:00pm each day(2/4)
Results
Fluorescence microscope to observe the GFP in leave
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 205
Illegal NgoMIV site found at 220
Illegal NgoMIV site found at 1567
Illegal NgoMIV site found at 1828
Illegal NgoMIV site found at 1990 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 4079
References
Wu, Y., Lei, C. F., Yi, D., Liu, P. M., & Gao, M. Y. (2011). Novel Bacillus thuringiensis δ-endotoxin active against Locusta migratoria manilensis. Applied and environmental microbiology, 77(10), 3227-3233.
| None |