This invention provides a
research model and construction method for
calcium ion imaging of ASH neurons in nematodes, belonging to the field of biological model construction technology. The construction method includes the following steps:
nematode culture; construction of a transgenic probe
plasmid: obtaining the ASH
neuron-specific
promoter sra-6 using PCR technology, and connecting
HindIII and
BamHI restriction sites to both ends of the
promoter; amplifying YC3.60 using PCR technology; then inserting the
promoter sequence into the pPD95.75
plasmid, and then inserting YC3.60 after the promoter. Transgenic
microinjection of nematodes is used, followed by screening to obtain nematodes with ASH neurons carrying
green fluorescent protein. In this application, YC3.60 is transferred into nematodes via transgenic means. Using a promoter specifically expressed in ASH neurons to link the
gene of this fluorescent probe
protein into the
nematode, the fluorescent probe can be specifically expressed in the
neuron. The constructed
nematode model can be used for neuronal structure and
functional imaging studies, as well as neuropharmacological
efficacy analysis.