Skip to main content
. 1997 Nov 25;94(24):13128–13133. doi: 10.1073/pnas.94.24.13128

Figure 3.

Figure 3

mec-4(d) vectors for transcriptional and translational gene fusions. pABL1, pABL2, and pABL3 enable translational fusions in any reading frame to be constructed. They differ in the nucleotides corresponding to the BclI site in pABL1; the BclI site is present only in pABL1 (this site is blocked by dam methylase and can only be used when DNA is prepared from a dam− host). Unique restriction sites that facilitate cloning are indicated. The mec-4(d) genomic sequence includes a conserved poly(A) addition site. The unc-54 3′ end cassette (24) is positioned after this site. Note that the unique Sse8387I site in the polylinker enables fragments with PstI ends to be inserted. In general, previously constructed lacZ and GFP fusion constructs made in standard C. elegans vectors (24, 28) can be readily converted to mec-4(d) fusion constructs. Swapping XbaI–ApaI or XbaI–SplI reporter + 3′ end cassettes might be the most common conversion strategy; for constructs without the nuclear localization signal, substituting an XbaI–ApaI fragment from pABL2 into the AgeI–ApaI region of the fusion construct should produce an in-frame mec-4(d) fusion equivalent to the original lacZ or GFP fusion.