Chen C L, Chang L K, Chang Y S, Liu S T, Tschen J S
Institute of Botany, National Chung-Hsing University, Taichung, Taiwan.
Mol Gen Genet. 1995 Jul 28;248(2):121-5. doi: 10.1007/BF02190792.
A total of 20 Bacillus subtilis F29-3 mutants defective in fengycin biosynthesis was obtained by Tn917 mutagenesis. Cloning and mapping results showed that the transposon in these mutants was inserted in eleven different locations on the chromosome. We were able to use the chromosomal sequence adjacent to the transposon as a probe to screen for cosmid clones containing the fengycin biosynthesis genes. One of the clones obtained, pFC660, was 46 kb long. Eight transposon insertion sites were mapped within this plasmid. Among the eleven different mutants analyzed, four mutants had Tn917 inserted in regions which encoded peptide sequences similar to part of gramicidin S synthetase, surfactin synthetase, and tyrocidine synthetase. Our results suggest that fengycin is synthesized nonribosomally by the multienzyme thiotemplate mechanism.
通过Tn917诱变获得了20株丰原素生物合成缺陷的枯草芽孢杆菌F29-3突变体。克隆和定位结果表明，这些突变体中的转座子插入到了染色体上11个不同的位置。我们能够使用转座子附近的染色体序列作为探针，筛选含有丰原素生物合成基因的黏粒克隆。获得的一个克隆pFC660长46 kb。在该质粒内定位了8个转座子插入位点。在分析的11个不同突变体中，有4个突变体的Tn917插入到了编码与短杆菌肽S合成酶、表面活性素合成酶和短杆菌酪肽合成酶部分相似的肽序列的区域。我们的结果表明，丰原素是通过多酶硫模板机制非核糖体合成的。
