Method for chimeric labeling of zebra fish primary motor neuron by fluorescent fusion protein

Abstract

The invention provides a method for chimeric labeling of zebra fish primary motor neurons by using fluorescent fusion protein, and relates to the technical field of transgenosis, and an establishment method of a system for chimeric labeling of zebra fish primary motor neurons by using fluorescent fusion protein comprises the following steps: by using donor plasmids p5E-mnx1, pME-EGFP-alpha-Tubulin and p3E-polyA and a skeleton plasmid pDestTol2pA2, carrying out fluorescent labeling on zebra fish primary motor neurons by using fluorescent fusion protein; the method comprises the following steps: obtaining a tissue specific expression plasmid pDest-mnx1-EGFP-alpha-Tubulin by virtue of an LR recombinant reaction, and carrying out transformation and plasmid extraction on the tissue specific expression plasmid pDest-mnx1-EGFP-alpha-Tubulin; a DNA fragment mnx1-EGFP-alpha-Tubulin is inserted into a part of cell genomes of zebrafish with AB genetic background through Tol2 transposase, a donor plasmid p5E-mnx1 is shown as a sequence 1 in a sequence table, EGFP is shown as a sequence 2 in the sequence table, and alpha-Tubulin is shown as a sequence 3 in the sequence table. A zebra fish motor neuron system marked by the microtubule-fluorescent fusion protein chimeric marker is established from a brand new angle, and clear marking of a single motor neuron is achieved. And an intuitive and rapid model is provided for exploring the development and morphogenesis of the motor neurons.

Description

technical field [0001] The invention relates to the field of transgenic technology, in particular to a method for chimerically marking zebrafish primary motor neurons with a fluorescent fusion protein. Background technique [0002] Motor neurons are one of the main cells that make up the nervous system, and their development depends on a variety of complex physiological and biochemical pathways to ensure the normal differentiation, maturation and connection of neurons. Under normal physiological conditions, motor neurons can form complex neural pathways in the body through structures such as axons and neuromuscular junctions, transmit nerve impulses from the central nervous system to various parts of the body, and regulate muscle activities, including breathing, walking and other delicate functions. motor skills. Apoptosis, degeneration, injury of motor neurons or slowing down of axonal neurotransmitter transmission can lead to neurodegenerative diseases such as motor neuro...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the technical problem that the transgenic zebrafish with primary motoneurons still has partial defects in the prior art

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