| Autor: |
Yu Xuping, Zhu Junhi, Yao Xunping, He Shicheng, Huang Haining, Chen Weiliang, Li Debao |
| Předmět: |
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| Zdroj: |
Chinese Science Bulletin; Jul2004, Vol. 49 Issue 13, p1370-1375, 6p, 1 Black and White Photograph, 4 Diagrams |
| Abstrakt: |
To understand the antagonistic mechanism of the broad spectrum antagonistic Enterobacter cloacae B8, TnS transposon-mediated mutagenesis is performed using suicide plasmid pZJ25. Two mutant strains that lost antagonistic character are isolated. Tagging with kanr gene on Tn5, an antagonistic related DNA fragment, the F fragment, right of the TnS insertion site is cloned in a plasmid named pTLF, from one of the mutant strains B8F. The 735 bp F fragment is then sequenced after subcloning. Genomic DNA of the original B8 strain is isolated, digested with Pst I and ligated to Pst I cassette. DNA fragments left and right of the F fragment are amplified from the Pst I cassette library using cassette primer and specific primers designed according to known sequence. 1106 bp sequence left of the F fragment and 664bp sequence right of the F fragment are finally obtained. Bioinformatics analysis shows that the contig assembled from the sequences of the cloned antagonistic related DNA fragments of B8 encodes three ORFs and is homogeneous to admM, admN and admO genes of Pantoea agglomerans andrimid biosynthetic gene cluster (AY192 157). The ORF, named anrF gene which encodes a polyketide synthase, knocked out by TnS insertion, is a homology of admM and the insertion site of Tn5 is at 214 bp upstream of the stop codon. It is concluded that the anrF gene is a gene related to the antagonistic activity of E. cloacae B8, and speculated that the antagonistic substance produced by B8 is an andrimid. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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