Difference between revisions of "Part:BBa K3895012"

Revision as of 17:26, 10 October 2021

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.

Figure 1. Plasmid construction of 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

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)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE 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
1000
INCOMPATIBLE 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.

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