Worm Breeder's Gazette 9(3): 78
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.
We have put a short sup-7 containing fragment into a variety of places in puc-family vectors to aid in constructions. The original plasmid, pAst contained a 1.4kb SalI - EcoRI fragment around the sup-7 gene inserted into puc-12 using the homologous sites. Plasmids pAst 18a, pAst18b, pAst 19a, and pAst 19b contain a 400 base HincII - RsaI fragment inserted in the SspI site of puc 18 and 19 respectively. The resulting plasmids should have all of the sites in the polylinker available for cloning (except possibly Acc1, SphI), and can still be used with the blue/white selection. The four plasmids have been checked for trainsient suppression of tra-3(am) by distal tip injection and all function well. We have not yet used these to make stably transformed lines, but foresee no problems. (So far all constructs that function in the transient expression assay have also been able to stably transform lines.) The pICt H and pICt R vectors incorporate a similar 400 b sup-7 containing fragment (this time SalI-RSAI) inserted between the SalI and EcoRV sites of pIC R and pIC H. These vectors can serve as a ready source of sup-7 containing restriction fragments either by cutting with RI or HinIII, by cutting with pairs of enzymes, or by excising and circularizing the RI or HinIII inserts and then cutting with any single enzyme in the polylinker.