Worm Breeder's Gazette 9(3): 78

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sup-7 Transformation Vectors

R. Waterston and A. Fire

Figure 1

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.

Figure 1