Aventis Pharma S.a.

Libragen

Centre National De La Recherche Scientifique

Cnrs

Karine Tuphile

Renaud Nalin

Benjamin Gillet

Pierre Monsan

Patricia Lacroix

Patrick Robe

Jean-Luc Pernodet

Michel M Guerineau

Claude Gerbaud

Pascal Simonet

Nathalie Oestreicher-Mermet-Bouvier

Sophie Courtois

Camela Cappellano

François Francou

Alain Raynal

Maria Ball

Guennadi Sezonov

Asa Frostegard

Pascale Jeannin

Pernodet Jean-Luc

Fatma Karray

Anne Gondran

Emmanuelle Darbon-Rongere

Helene Dominguez

Marie-Helene Blondelet-Rouault

Pierre Pujic

Philippe Reulet

Claude Bensoussan

Petar Pujic

Nathalie Oestreicher-Mermet- Bouvier

Hélène Dominguez

Marie-Hélène Blondelet-Rouault

The present invention relates to methods for the identification or cloning of polynucleotides encoding a selected phenotype, particularly from environmental DNA. In a specific embodiment, the method comprises (i) cloning environmental DNA fragments intocloning vectors to produce a metagenomic library, (ii) identifying or selecting cloning vectors in the library which contain DNA fragments having a particular characteristic of interest, (iii) modifying the identified or selected cloning vectors into shuttle or expression vectors for transfer and integration in a selected host cell, (iv) transferring the modified cloning vectors into the selected host-cell and (v) identifying or cloning the DNA fragments contained in the modified cloning vectors which encode the selected phenotype in the selected host cell.

Method for the expression of unknown environmental DNA into adapted host cells

The invention relates to the direct selection of metabolic pathways having a determined function in the transformation of a substrate {Ai} into a target product {B}, which is of interest in the industrial, pharmaceutical or agri-food sectors. More specifically, the invention relates to the detection, within metagenomic libraries, of novel biosynthesis pathways involved in a biochemical reaction having a known product {B}. The selection and characterization of said novel metabolic pathways enables {B} to be produced enzymatically. The invention provides an alternative to the chemical synthesis of the molecule in question {B}. Moreover, and above all, the invention can be used specifically to target and exploit the only metabolic pathways enabling the transformation of {Ai} into {B}, while eliminating the associated metabolic pathways that can catabolise the target product {B}.

Method for the identification of a metabolic pathway family by means of positive selection

The present invention relates to the isolation and identification of novel genes of the biosynthetic pathway for spiramycins and to novel polypeptides involved in this biosynthesis. The invention also relates to a method for producing these polypeptides. It also relates to the use of these genes for the purpose of increasing the levels of production and the purity of the spiramycin produced. The invention relates in particular to a microorganism which produces spiramycin I but which does not produce spiramycin II and III, and to the use of such a microorganism. The invention also relates to the use of the genes of the biosynthetic pathway for spiramycins for constructing mutants which can lead to the synthesis of novel antibiotics or to derived forms of spiramycins. The invention also relates to the molecules produced through the expression of these genes and to pharmacologically active compositions of a molecule produced through the expression of such genes.

Polypeptides involved in the biosynthesis of spiramycins, nucleotide sequences encoding these polypeptides and applications thereof

The invention concerns a method for preparing nucleic acids from an environment sample, more particularly a method for obtaining a library of nucleic acids from a sample. The invention also concerns nucleic acids of nucleic acid libraries obtained by said method their use in the synthesis of novel compounds, in particular novel compounds of therapeutic interest. The invent further concerns novel means used in the method for obtaining said nucleic acids, such as novel vectors and novel processes for preparing such vectors or recombinant host cells containing said nucleic acid. Finally, the invention concerns methods for detecting a nucleic acid of interest within a library of nucleic acids resulting from said method, and nucleic acids detected by said method and polypeptides encoded by said nucleic acids.

Method for obtaining nucleic acids from an environment sample, resulting nucleic acids and use in synthesis of novel compounds

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Caluire, France

Karine Tuphile