CONTROLLABLE ON-OFF METHOD FOR FISH REPRODUCTION

摘要

The invention discloses a method for controlling fish reproduction, the method comprises the following steps of: A, constructing a recombinant gene CMV-eGFP-SV40-CMV-Ga14-SV40 and establishing a GAL4 transgenic zebrafish line; B, constructing a recombinant gene CMV-RFP-SV40-UAS-antisense dnd and establishing a uas-antisensednd transgenic zebrafish line; and C, hybridizing by taking homozygous transgenic fish of the two lines as parents, and the hybrid generation fish was sterility. The method of the present invention takes a strategy that two fertilizable lines hybridized but hybrid generation is sterility, establishing a novel method with a universal meaning for controlling fish fertility; The method solves the contradiction of the reproductively sterility of fish and the hereditability of characters in a reproduction operation research and can be effectively applied to cultivation of new fish specie and population control.

主权项

1. A method of controlling fish reproduction, characterized in that, comprising the following steps:
(A) Introducing exogenous genes: introducing recombinant genes CMV-eGFP-SV40-CMV-Gal4-SV40 and CMV-RFP-SV40-UAS-antisense dnd into fertilized eggs of zebrafish by microinjection method to prepare P0 generation of GAL4 transgenic zebrafish and UAS-antisense dnd transgenic zebrafish; The recombinant gene CMV-eGFP-SV40-CMV-Gal4-SV40 has a sequence shown in SEQ ID NO: 1; The recombinant gene CMV-RFP-SV40-UAS-antisense dnd has a sequence shown in SEQ ID NO: 2; (2) Obtaining homozygous transgenic fish: fish that expressing green fluorescent protein is P0 generation GAL4 transgenic zebrafish; and fish that expressing red fluorescent protein is P0 generation UAS-antisense dnd transgenic zebrafish; hybridizing the two P0 transgenic zebrafish with control zebrafish, respectively, to obtain F1 generation GAL4 transgenic zebrafish and UAS-antisense dnd transgenic zebrafish; inbreeding the F1 generation GAL4 transgenic zebrafish, to obtain homozygous GAL4 transgenic zebrafish in F2 generation; and inbreeding the F1 generation UAS-antisense dnd transgenic zebrafish to obtain homozygous UAS-antisense dnd transgenic zebrafish in F2 generation; (3) Hybridizing transgenic fish: hybridizing, by taking the homozygous GAL4 transgenic zebrafish and homozygous UAS-antisense dnd transgenic zebrafish as parents, the transgenic fish of two lines, to produce sterilized fish offspring.