If you are experiencing problems downloading PDF or HTML fulltext, our helpdesk recommend clearing your browser cache and trying again. If you need help in clearing your cache, please click here . Still need help? Email help@ingentaconnect.com

Sequencing and modular analysis of the hybrid non-ribosomal peptide synthase - polyketide synthase gene cluster from the marine sponge Hymeniacidon perleve-associated bacterium Pseudoalteromonas sp. strain NJ631

$50.00 plus tax (Refund Policy)

Buy Article:

Abstract:

In the present study, we sought to confirm a putative non-ribosomal peptide synthase (NRPS) - polyketide synthase (PKS) gene cluster in marine sponge-associated bacterium with cytotoxic activity and elucidate the gene’s structural information. The genomic library of the marine sponge Hymeniacidon perleve-associated bacterium Pseudoalteromonas sp. strain NJ631 was constructed using a pCC1FOS fosmid. Positive clones that covered the whole gene cluster region of hybrid NRPS-PKS were selected for shotgun sequencing. The results obtained from BlastX and open reading frame (ORF) analysis indicated that there are 3 big ORFs, NJA1, NJA2, and NJA3, that encoded proteins with similarities to amino acid adenylation, -ketoacyl synthase, and non-ribosomal peptide synthase, respectively, from different organisms. The results gave us a clue that there could be PKS or NRPS modules in the 3 ORFs. Further analysis demonstrated 3 ORFs encoding 2 NRPS modules, 1 PKS module, and 3 NRPS modules. Using the specificity-conferring selection rule, the substrate specificity of 4 adenylation (A) domains (A2, A3, A4, and A5) were successfully predicted, and the amino acids of the substrate specificity were glutamic acid - glutamine, serine, d-serine, and Aeo (2-amino-9,10-epoxy-8-oxodecanoic acid), respectively.

Nous tentons de confirmer dans cette étude la présence d’une grappe de gènes des complexes NRPS-PKS chez des bactéries associées à des éponges marines possédant une activité cytotoxique, et d’en élucider la structure. Une banque génomique de la bactérie Pseudoalteromonas sp. NJ631 associée à l’éponge marine Hymeniacidon perleve a été construite avec le fosmide pCC1FOS. Les clones positifs qui couvrent la région complète de la grappe de l’hybride NRPS-PKS ont été choisis pour un séquençage à l’aveugle. Les résultats obtenus des analyses BlastX et des cadres de lectures ouverts ont indiqué qu’il existe 3 grands cadres de lecture ouverts, NJA1, NJA2 et NJA3 qui codent des protéines possédant des similarités avec l’adénylation d’acides aminés, des -cétoacyl synthétase et de peptides synthétases non ribosomales, respectivement, de différents organismes. Ces résultats nous donnent un indice à l’effet qu’il pourrait exister des modules PKS ou NRPS dans ces trois cadres de lecture ouverts. Une analyse supplémentaire démontre que les trois cadres de lecture ouverts codent deux modules NRPS, 1 module PKS et 3 modules NRP. Grâce à la règle de sélection conférant la spécificité, la spécificité de substrat des 4 domaines (A2, A3, A4, A5) a été prédite avec succès et les acides aminés qui confèrent cette spécificité sont l’acide glutamique - le glutamine, la sérine, la d-serine et l’Aeo.

Document Type: Research Article

Publication date: March 1, 2009

More about this publication?
  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
  • Information for Authors
  • Submit a Paper
  • Subscribe to this Title
  • Terms & Conditions
  • Sample Issue
  • Reprints & Permissions
  • ingentaconnect is not responsible for the content or availability of external websites
Related content

Tools

Favourites

Share Content

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more