Part:BBa_K4752008
GLR2.9-Bx reporter
Description
The GLR2.9 promoter is a specific regulatory region within the plant genome that becomes active in response to attacks by pests and diseases. When plants are threatened by pests or diseases, the GLR2.9 promoter is triggered into activation, initiating a biological response. This activation leads to the expression of the Bx reporter system.
Bx functions as a reporter system, manifesting a noticeable yellow coloration in the leaves of the plant when activated by the GLR2.9 promoter. The change in leaf color serves as a visible signal, allowing for easy identification of plants facing pest or disease stress. This feature is highly valuable for monitoring and researching plant responses to such stresses, aiding in the understanding and management of plant health.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 630
Illegal EcoRI site found at 1257
Illegal XbaI site found at 1600
Illegal PstI site found at 3907 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 630
Illegal EcoRI site found at 1257
Illegal PstI site found at 3907 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 630
Illegal EcoRI site found at 1257
Illegal BglII site found at 1067
Illegal BglII site found at 1675
Illegal BglII site found at 2293 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 630
Illegal EcoRI site found at 1257
Illegal XbaI site found at 1600
Illegal PstI site found at 3907 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 630
Illegal EcoRI site found at 1257
Illegal XbaI site found at 1600
Illegal PstI site found at 3907 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5
Illegal SapI.rc site found at 2030
Illegal SapI.rc site found at 3560
Assembly
The NcoI-PstI double digestion was performed on the pCAMBIA1300-C-MCS-GFP plasmid to obtain a linearized vector fragment. Next, homologous recombination was carried out by ligating the GLR2.9 promoter (BBa_K4752004) DNA fragment, Bx reporter gene fragment, and the linearized vector fragment, resulting in the expression vector plasmid containing the GLR2.9-Bx reporter system. These plasmids were then transformed into DH5α Escherichia coli and spread on LB agar plates supplemented with kanamycin for selection. Single colonies were picked from the plates and subjected to PCR and agarose gel electrophoresis verification, followed by sequencing to ensure correct plasmid construction.
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