Worm Breeder's Gazette 17(2): 19 (April 1, 2002)

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

New and improved pie-1-based vectors for maternal expression of transgenes

Ingrid D'Agostino, Kimberly Reese, Geraldine Seydoux

Dept. of Molecular Biology and Genetics, Johns Hopkins U. School of Medicine725 North Wolfe Street / 515 PCTB, Baltimore MD 21205

We previously reported the construction of a pie-1-based vector for maternal
 expression of GFP fusion proteins in the germline and early embryos [Worm
 Breeder's Gazette 15(5): 18 (February 1, 1999)]. This vector was modified by
 Praitis et al., 2001 for use in bombardment-mediated transformation. We have now
 created new versions of these vectors that are compatible with Invitrogen's
 Gateway Recombination Cloning Technology. This technology bypasses the need for
 restriction enzymes and allows for the directional cloning of virtually any ORF
 into the pie-1 vectors. 

For expressing ORFs in the germline: 

unc-119 rescuing fragment/pie-1 promoter-gateway destination cassette B-3'utr of pie-1 
(suitable for bombardment, Praitis et al., 2001)

For expressing GFP-ORF fusions in the germline:

pie-1 promoter-GFP- gateway destination cassette B -3'utr of pie-1
(suitable for complex arrays, Kelly et al., 1997)

unc-119 rescuing fragment / pie-1 promoter-GFP- gateway destination cassette B - 3'utr of pie-1
(suitable for bombardment, Praitis et al., 2001)

Anyone interested in these vectors should write to Geraldine at gseydoux@jhmi.edu.