Study on transgenic efficiency of Fst1 gene element mediated by Tgf2 transposon in common carp

Abstract:

Follistatin (FST) could antagonize the TGF-β superfamily members and inhibit the activity of endo-myostatin via direct protein-protein interaction, thereby restoring muscle growth. Here, according to the splicing results of the high-throughput transcriptome sequencing data for Cyprinus carpio, we cloned the open reading frame (ORF) sequence of the Fst1 gene, which was 1 260 bp in length and encodes a protein of 320 amino acids. We then constructed a pTgf2-Mylz2-ccfst1 donor plasmid carrying both the left (220 bp) and right (185 bp) arms of goldfish Tgf2 transposon, the promoter of zebrafish Mylz2 and the ORF of carp Fst1. Transgenic carps were obtained via co-microinjection of the donor and the capped mRNA of the Tgf2 transposase synthesized in vitro. The average integration rate of the exogenous gene in transgenic carp was 44.7% as measured by PCR. The amplified products of four positive transgenic carp were recovered, cloned and sequenced to confirm that all of them contained the target fragment. Our results demonstrated the high transgenic efficiency of the Tgf2 transposon in C. carpio, which laid the foundation for further studies on the function of FST during carp muscle development.