Identification of DVA Interneuron Regulatory Sequences in Caenorhabditis elegans
Figure 5
Mutation analysis. A. Analysis of the twk-16 intron and enhancer.
Deletion analysis of the twk-16 intron with 73 bp twk-16.cs1 (cs1) and 259 bp twk-16.cs2 (cs2) denoted in red with flanking sequences in black and not to scale. The approximate sizes of the wild-type (WT) sequences are denoted by numbers from WT2000 to WT53. WT2000 was a plasmid construct and contains 500 bp 5′ of exon 1, exon 1 and 1.4 kb of the first intron containing both cs1 and cs2. WT700 contains 38 bp of flanking sequences 5′ to cs1 and 244 bp of flanking sequences 5′ to cs2 and 102 bp of 3′ flanking sequences. WT350 contains 55 bp of flanking sequences 5′ to cs2 and 45 bp of 3′ flanking sequences. WT500 contains 202 bp of flanking sequences 5′ to cs1 and 223 bp of 3′ flanking sequences. W300 contains 114 bp of flanking sequences 5′ to cs1 and 121 bp of 3′ flanking sequences. WT195 contains 114 bp of 5′ flanking sequence to cs1 and 8 bp of 3′ flanking sequence. WT113 contains 23 bp of 5′ flanking sequences to cs1 and 17 bp of 3′ flanking sequence. WT85 contains 4 bp of 5′ flanking sequences to cs1 and 8 bp of 3′ flanking sequence. WT85 contains WT53 with 17 bp of 5′ flanking sequence and 15 bp of 3′ flanking sequence. WT53 contains 53 bp of the 73 bp twk-16.cs1 region. B. Mutational analysis of WT53. The wild-type sequence is denoted by black type with the mutations of WT53 shown in red type. Conserved sequences identified by the seven species MUSSA comparison are in blue type with blue underlining. WT29 and WT24 are generated by cleavage of WT53 within the Mut3 region. DVA or broad neuronal expression is denoted by+or – in the box to the right of each construct.