Journal:

Article Title: Cytochrome c 3 Mutants of Desulfovibrio desulfuricans

doi:

Figure Lengend Snippet: Diagrammatic representation of the D. desulfuricans G20 chromosome region encoding cycA. (A) Arrangement of genes in wild-type genomic DNA. The vertical arrow indicates the position of a putative promoter. The black bar indicates the position of a putative ORF upstream of cycA that is transcribed left to right, as is cycA in this illustration. t1 and t2 indicate the locations of possible transcriptional terminators. (B) Plasmid pBSC2 (Table ​(Table1)1) used for gene disruption. The SacII-PstI internal cycA fragment is designated ‘cycA’. (C) Plasmid-interrupted chromosomal cycA. The positions of PCR and sequencing primers, B1 cyc primer and Km primer for the leftward junction and T3 primer and cyc2 left primer for the rightward junction, are indicated by arrows. The left copy of cycA lacks 63 bp of the 3′ end of the coding sequence (which totals 393 bp), while the right copy lacks 71 bp of the 5′ end. Note the different scale for panel A versus that for panels B and C.

Article Snippet: When a PCR product was not obtained with these primers, the DNA from the putative mutant was digested with Sac II (Fig. C) and recircularized to generate a Km r Ap r plasmid, and the cycA gene was sequenced from the T3 primer (Promega).

Techniques: Plasmid Preparation, Sequencing