Cloning and functional analysis of the TERE1 gene using the Gal4-UaS system in S2 cells: A streamlined approach for human gene functional genomics

Abstract

The post-genome sequencing era faces the challenge of understanding gene functions. While model organisms and mutagenesis screens are helpful, they can be labor-intensive. This study demonstrates a streamlined gene cloning approach using the human TERE1 gene and the pUAST vector system for expression in Drosophila S2 cells. The pUAST vector was employed to clone the TERE1 gene and insert it into S2 cells, allowing co-transfection with multiple constructs for co-expression of up to four proteins. This provided a stable and selectable platform. The cloning of TERE1 in PcDNA3.1 and pUAST vectors confirmed the expression of TERE1 protein alongside EGFP. The expressed TERE1 protein in Drosophila melanogaster (D.m) Schneider recombinants resembled the normal Drosophila HEIX1 protein, which is critical in the larval-to-adult transformation as proved by western blot. The pUAST system proved effective for TERE1 gene cloning and the simultaneous expression of multiple proteins. This simplified method presents an efficient alternative to traditional mutagenesis screens, facilitating gene function studies and aiding the identification of disease-related genes.