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Cloning and Identification of Genes Involved in Biosynthesis of Antagonistic Substance from Bacillus Subtilis Strain B11

Author: WuXue
Tutor: FengJiaXun
School: Guangxi University
Course: Biochemistry and Molecular Biology
Keywords: Bacillus subtilis genomic library antagonism polyketide synthase gene cluster
CLC: S476
Type: Master's thesis
Year: 2006
Downloads: 31
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Abstract


Bacillus subtilis strain B11 displayed antagonistic activity against a wide variety of plant pathogenic fungi and bacteria. A genomic library of the strain B11 containing 9000 clones was constructed. The average size of the inserted DNA in recombinant plasmids was 42.1 kb and the probability of harboring any gene in the genome of the strain B11 was 99.99%. The library was screened for antagonistic activity against Ralstonia solanacearum strain P13 and a positive clone carrying a 50 kb DNA insert was isolated, which was designated as GXN9527. Plate assay showed that GXN9527 could also inhibit the growth of plant pathogenic Gram-negative bacterium Xanthomonas oryzae pv. oryzae, but not inhibit the Gram-positive bacterium Bacillus licheniformis or the phytopathogenic fungi Fusarium oxysporum f.sp.niveum, and Rhizoctonia solani. pGXN9527 was subcloned and partially sequenced. A continuous DNA sequences with 28.481 kb was assembled. The 28.481 kb DNA sequence contained a PKS gene cluster(pksA～pksF) that shared 96%～99% homology with the pks2 gene cluster from Bacillus amyloliquefaciens strain FZB42. The pksA had a 2028 bp ORF encoding a product with 675 amino acids sharing 96% identities and 97% positives with Pks2D of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAG23966.1); The pksB had a 6999 bp ORF encoding a product with 2332 amino acids sharing 96% identities and 98% positives with Pks2E of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAG23967.1); The pksC had a 5721 bp ORF encoding a product with 1906 amino acids sharing 97% identities and 98% positives with Pks2F of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAG23968.1); The pksD had a 7386 bp ORF encoding a product with 2441 amino acids sharing 97% identities and 98% positives with Pks2G of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAG23969.1); The pksE had a 3855 bp ORF encoding a product with 1284 amino acids sharing 96% identities and 97% positives with Pks2H of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAG23970.2); The pksF had an 1128 bp ORF encoding a product with 375 amino acids sharing 97% identities and 98% positives with Pks21 of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAJ57404.1); The pdhG had an 871 bp ORF encoding a product with 290 amino acids sharing 98% identities and 99% positives with PdhA of Bacillus amyloliquefaciens FZB42 (GenBank accession No. CAJ57405.1).

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control
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