TRANSPOSON MUTAGENESIS AND CLONING OF THE GENES ENCODING THE ENZYMES OF FENGYCIN BIOSYNTHESIS IN BACILLUS-SUBTILIS

被引:33
作者
CHEN, CL
CHANG, LK
CHANG, YS
LIU, ST
TSCHEN, JSM
机构
[1] CHANG GUNG COLL MED & TECHNOL,DEPT MICROBIOL & IMMUNOL,GENET MOLEC LAB,TAYUAN 333,TAIWAN
[2] NATL CHUNGHSING UNIV,INST BOT,TAICHUNG 402,TAIWAN
来源
MOLECULAR & GENERAL GENETICS | 1995年 / 248卷 / 02期
关键词
FENGYCIN; ANTIFUNGAL ANTIBIOTIC; LIPOPEPTIDE ANTIBIOTIC; NONRIBOSOMAL MULTIENZYME THIOTEMPLATE MECHANISM; BACILLUS SUBTILIS;
D O I
10.1007/BF02190792
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页码:121 / 125
页数:5
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