Issue Information

Abstract

Current ProtocolsVolume 3, Issue 2 e467 ISSUE INFORMATIONFree Access Issue Information First published: 25 February 2023 https://doi.org/10.1002/cpz1.467AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Graphical Abstract Cover: Synthetic assembly DNA cloning for Drosophila melanogaster is described in Venken et al. Different DNA parts, obtained from flies directly, other species, or through synthetic means, are put together using synthetic assembly DNA cloning to generate continuously growing plasmids of increasing complexity. The resulting end products can be two transgenes, a first transgene providing G418 sulfate selection (provided by the NeoR marker) and encoding the binary LexA transactivator driven by the R70B04 enhancer (i.e., the G418 sulfate-selectable LexA transactivator plasmid), and a second transgene providing Blasticidin S selection (provided by the BlastR marker) and encoding the binary LexA-Op responder, reporting green fluorescent protein (sfGFP) reporter expression driven by the LexA binary transactivator, whose expression is regulated by the R70B04 enhancer (i.e., the Blastidicin S-selectable LexA-Op responder plasmid). A double transgenic fly can then be obtained directly by injecting both transgenes (i.e., dual transgenesis) followed by co-selection using both G418 sulfate and Blasticidin S. The double transgenic fly can then be analyzed for expression patterns. Expression analysis of dual transgenic larval animals shows green fluorescent protein (sfGFP) reporter expression against the neuropil marker Discs Large (Dlg) in the larval central nervous system. View all latest articles RelatedInformation