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Resources » Paper

Chris Merritt et al. (2007) International Worm Meeting "A Modular Vector Kit For Transgenesis in C. elegans."

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    Publication type:
    Meeting_abstract
    WormBase ID:
    WBPaper00030472

    Chris Merritt, Dominique Rasoloson, Christopher Gallo, Ekaterina Voronina, Darae Ko, & Geraldine Seydoux (2007). A Modular Vector Kit For Transgenesis in C. elegans presented in International Worm Meeting. Unpublished information; cite only with author permission.

    We have constructed a new vector kit for transgenic expression in somatic and germline tissues of C. elegans. This kit was created to facilitate the cloning of promoter, ORF and 3UTR sequences in any desired combination. By allowing users to choose both promoters and 3UTR sequences, this kit can be used for genes expressed in the soma (promoter-dependent) as well as genes expressed in the germline (3 UTR-dependent, see abstract by Merritt et al.). The vector kit is based on MultiSite GATEWAY recombination cloning technology, which permits simultaneous ligation of up to three DNA fragments in one step. The kit consist of 1) a destination vector containing the unc-119 gene for selection in biolistic transformation, and 2) three sets of donor clones containing either promoter sequences (heat shock promoter for inducible expression and pie-1 promoter for germline expression), ORFs (including GFP:actin, GFP:tubulin, GFP:H2B and GFP), or 3UTR sequences (including unc-54 and several germline 3UTRs). The kit is also compatible with the Vidal/Hope collections of promoters and ORFs. The modular vector kit can be used 1) to define gene expression patterns, 2) to express ORFs ectopically and 3) to define the contributions of promoter, ORF and 3UTR sequences in specifying expression patterns. For example, we are currently using this system to systematically characterize the role of 3UTRs in specifying expression patterns in the germline. Using a generic germline promoter (pie-1) and a generic ORF (GFP:H2B), we have found that many germline expression patterns are specified by 3UTR sequences (see abstract by Merritt et al.).


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