Generation and Confirmation of Human TAL2 Transgenic Drosophila Model

Abstract

TAL2 is a transcription factor of the basic helix-loop-helix (bHLH) family. TAL2 may be an oncogene that is overexpressed as a consequence of the chromosomal translocation in T-cell acute lymphoblastic leukemia. Recent studies have indicated that TAL2 involved in development of many tumors. Although aberrant TAL2 expression was associated with tumorigenicity, TAL2 has been shown to play a crucial role in normal brain development. However, its role in neural development is unknown. In this study, molecular cloning and microinjection technology were used to construct UAS-TAL2 transgenic Drosophila model. The coding sequence of human TAL2 gene was achieved by gene synthetic technology and subcloned into pUAST expression vector. The pUAS-TAL2 recombinant vector was microinjected into the embryos of wild-type flies. The UAS-TAL2 transgenic flies with red eyes were screened out with mini-white marker, then the balancing and mapping of these strains were conducted. The integration of transgene into fly genome was confirmed through PCR amplification and the expression of TAL2 gene was examined by quantitative PCR. The results showed that pUAS-TAL2 recombinant plasmid was successfully constructed, and four independent transgenic strains were obtained after the screening and balancing. PCR analysis showed that the P[mini-white, UAS-TAL2] had integrated into genomes of transgenic strains and could express under regulations. The expression level of TAL2 gene in adult brains was upregulated significantly under elav-GAL4 driver. In conclusion, the UAS-TAL2 transgenic Drosophila model has been successfully generated, which lays the foundation for further studies of the function and regulation of human TAL2 gene in the neural development with GAL4/UAS system.