Worm Breeder's Gazette 12(2): 17 (January 1, 1992)

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.

Vectors for ectopic expression in C. elegans

A. Fire[1], S. White-Harrison[2], D. Dixon[3]

Figure 1

[1]Carnegie Institution of Washington, 115 West University Parkway, Baltimore, Md 21210
[2]Carnegie Institution of Washington, 115 West University Parkway, Baltimore, Md 21210
Present Address: Tufts University, Sacler School of Biomedical Sciences

[3]Carnegie Institution of Washington, 115 West University Parkway, Baltimore, Md 21210
Present Address: University of British Columbia, Dpt of Microbiology.

We have constructed a set of expression vectors intended to allow production of RNA or protein products of interest in well defined cells and/or temporal patterns. Each vector contains upstream sequences from a regulated promoter joined to a polylinker just before the ATG of the gene. This should allow production of protein products starting from the ATG in their own mRNA or genomic sequence.

The general structure of the vectors is shown in the figure. The design is modular, with an abundance of restriction sites in defined positions to facilitate transfer of equivalent cassettes between vectors. The vectors are related to the modular lacZ fusion vectors (Gene 93: 189), so that cassettes can often be exchanged between the two vector sets.

The 'canonical' vector pPD49 .26is a promoter-less template which is essentially the starting point for the constructions. This vector has just two signals: a synthetic intron and the 3' region from the unc-54 gene. These signals are interspersed between three consecutive MCS (polylinker) segments. Vectors based on pPD49 .26containing a variety of regulated promoters in the 5' MCS have been constructed. These vectors are summarized in the table. Each vector being distributed has been tested by inserting the lacZ gene into the expression MCS, and examining the pattern of ß-Gal activity in transgenics (either as F1 following injection or in heritable transformed lines carrying arrays.)

We will maintain a 'kit' of expression vectors that includes the vectors, sequences, and documentation. Anyone interested in obtaining vectors should request them from A. Fire. We encourage other laboratories to use the template vector pPD49 .26to generate expression vectors with new specificities. In addition to the vectors shown in the table, any tested expression vector which is sent to us will be included in the kit. Other casettes with general value for marking cells (e.g. by causing cell death or novel enzyme staining, or directing intracellular localization) will be 'collected' along with the vector set.

[See Figure 1]