Cloning and function of sanJ-A gene involved in nikkomycins biosynthesis of Streptomyces ansochromogenes

A 7.5kb DNA fragment was identified from the cosmid library of Streptomyces ansochromogenesus using a partial DNA fragment involved in the nikkomycin biosynthesis as a probe. It was cloned into pBluescript M13- and the resulting recombinant plasmid was designated as pNL2200. The nucleotide sequence of 2.3kb Sal I -BamH I DNA fragment was determined and analysed. The result indicated that the fragment contains one complete open reading frame (ORF); The start codon is GTG at 271 position and the stop codon is TGA at 1 954 position. It contains 1 686 base pairs and encodes a protein with 561 amino acids. The deduced product has 44% identity with the ligase belong to the superfamily of adenylate-forming enzyme. The experiment of gene disruption showed that the sanJ gene was closely related to nikkomycins biosynthesis of Streptomyces ansochromogenes. It was designated as sanJ.