The experiments were based on the basic reaction
for GABA production by
Dimocarpus longan Lour.cells.
To increase the GABA content of longan
cells, we based our experimental
design on two genes.
①.Knockdown of the CaM1 and CaM2 genes
The pGW3 (Genovo-w-3 system, Source: The New
Zealand Institute for Plant and Food Research Limited)
was constructed and two CaM genes native to
Longan cells: lab
numbers >Dlo025420 and >Dlo030022 were targeted for knockout.
Entry of pGW3 plasmid into recipient cells:
We obtained groups of Lobelia cells with deletion of all CaM genes.
②.Construct an overexpression vector for the GAD gene
We construct the GAD gene vector, and the GAD gene was delivered into
longan cells using
pCAMBIA-1301 plasmid, the GAD gene was amplified in the chromosome of longan cells, and the
GAD gene was doubly expressed.
By idealizing the genetic system of
longan itself through the above two
means, the output of
GABA is maximized.The pathways of GABA production in modified longan are as follows.
Meanwhile, based on bioinformatic
analysis of laboratory transcriptome
data, we found that
key genes for GABA production in longan respond to blue light and high temperature stress.
After transforming the original gene of longan, it was treated with exogenous treatment:
high temperature stress and blue light
High-temperature stress
Blue light treament
↓
Concept
