Construction and identification of UAS-Hey transgenic fly strains / 重庆医学

ABSTRACT

Objective To construct the UAS-Hey transgenic fly strains to provide a tool for researching the function of Hey gene in the fly development.Methods The Hey gene coding sequence was amplified by reverse transcription PCR and subcloned into pUAST expression vector.The pUAS-Hey recombinant plasmid was constructed and microinjected into the embryos of wild type flies.The UAS-Hey transgenic flies with red eyes were screened out with mini-white marker,then the balancing and mapping of these transgenic strains were performed.The identification was performed by using the PCR amplification method.Results The pUAS-Hey recombinant plasmid was successfully constructed,and seven independent transgenic strains were obtained by microinjection and transgenic fly screening and balance.PCR analysis confirmed that the P[mini-white,UAS-Hey] was integrated into genomes of transgenic strains and was in expressible region.Conclusion The UAS-Hey transgenic flies are successfully constructed,which lays the foundation for further studies of the function and regulation of Hey gene with GAL4/UAS system.