A method to obtain whole-kidney zebrafish expressing green fluorescent protein

Abstract

The invention discloses a method for obtaining a holonephros expression green fluorescent protein zebra fish. The method can overcome the defects that green fluorescent protein of an existing zebra fish line can not be evenly and simultaneously expressed in a whole original glomerulus, a tubule and a catheter. Green fluorescent protein of a zebra fish in the method can be simultaneously expressed in a whole original glomerulus, an original kidney tubule and an original kidney catheter. The method is a crossbreeding method according to which an Na, K-ATPase alpha1A4: GFP line zebra fish and a wt1b: GFP line zebra fish are mutual parents. The method for obtaining the holonephros expression green fluorescent protein zebra fish includes the following steps of hybridization parent selection and breeding, matching and breeding, hybridization character sieving, hybridization filial generation breeding and hybridization character fixing and retaining. The method for obtaining the holonephros expression green fluorescent protein zebra fish is a separation element hybridization stable integration method, the implementation process is easy to control, a zebra fish line of which green fluorescent protein can be simultaneously, evenly and efficiently expressed in a whole original glomerulus, an original kidney tubule and an original kidney catheter is obtained and the requirement for integrality and completeness of study on the original kidney functions of the zebra fish is met.

Description

technical field [0001] The invention relates to a zebrafish breeding method, in particular to a breeding method for obtaining whole kidneys expressing green fluorescent protein zebrafish, and belongs to the field of biotechnology. Background technique [0002] In 1962, Shimomura et al. first discovered the bioluminescent protein aequorin in Aequoria victoria, and observed a by-product of light green fluorescence under ultraviolet irradiation-Green Fluorescent Protein (Green Fluorescent Protein, GFP). In 1974, Shimomura et al. completed the purification of GFP, and Prasher et al. cloned its gene 18 years later. In 1994, Chalfie et al. realized the expression of GFP in E. coli cells and Caenorhabditis elegans. As an important biomarker protein, GFP can self-catalyze to form a chromophoric structure and emit green fluorescence under ultraviolet or blue light excitation. Because GFP has many advantages such as stable fluorescence, safety and non-toxicity, easy detection and ea...

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Problems solved by technology

In 2007, Yan Liu et al. produced the Na, K-ATPasealpha1A4:GFP transgenic strain zebrafish, and realized the specific expression of GFP in the zebrafish proto-renal tubule and pro-nephric duct, but in the pro-glomerulus of the zebrafish strain There is no specific green fluorescent expression in

At 35 hours after fertilization of the transgenic zebrafish line, green fluorescence can be observed in the pronephric ducts of the pronephric glomerulus and the proximal part, but no specific GFP expression can be observed in the distal ducts

When the above two strains of zebrafish are used for primary kidney research, GFP cannot be uniformly expressed in the entire primary glomerulus, tubules and ducts at the same time, which affects the integrity and integrity of the zebrafish primary kidney function research

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