动物实验,斑马鱼sat1.a z-bo 2020-09-29 387

　　Purpose: In the previous study, we screened a batch of tissue-specific zebrafish strains expressing green fluorescent protein GFP through insertion mutation mediated by Tol2 transposon. One of the strains, Tol2:20141221t, expresses GFP in the nervous system, but it has not been determined where the Tol2 transposon is inserted in the genome and which gene mutation is triggered. The main research goal of this paper is to identify zebrafish variants induced by the Tol2 transposon insertion.

　　Method: Use thermal asymmetric interface PCR (TAIL-PCR) to locate the genome inserted by the Tol2 transposon. In situ hybridization is used to detect whether the temporal and spatial expression of the mutant gene matches the GFP expression of the strain. Screen and identify homozygous variants, and further analyze the developmental defects caused by genetic mutations.

　　Results: In this strain, the Tol2 transposon was inserted into the 8th inclusion of spermidine/sperm N1-acetyltransferase 1a (spermidine/sperm N1-acetyltransferase 1a, sat1.a) In the subregions, the transcription of sat1 and .a genes was terminated prematurely. The homozygous variant of sat1.a was screened, but no obvious developmental disorders were detected.

　　Conclusion: The screened and identified Tol2 transposon-mediated zebrafish sat1.variant has no obvious developmental defects, but it can be used as a powerful tool to study the development of the nervous system.