Sinvent As

Vectron Biosolutions As

Fmc Biopolymer As

Leiv Eiriksson Nyfotek As

Algipharma As

Svein Valla

Trond Erling Ellingsen

Sergey Zotchev

Trygve Brautaset

Rahmi Lale

Olga Nikolayivna Sekurova

Ole-Martin Gulliksen

Arne Reidar Strøm

Håvard Sletta

Espen Fjærvik

Ingrid Bakke

Laila Berg

Gudmund Skjak-Braek

Havard Sletta

Espen Fjaervik

Martin Gimmestad

Karuna Ponniah Karunakaran

Karianne Bakkevig

Simone Balzer Le

Trond Erik Vee Aune

Helga Ertesvag

Hävard Sletta

Janet M Blatny

Ponniah Karunakaran

Trine Aakvik

Trond Ellingsen

Trond Erik Aune

Kristin Fløgstad Degnes

Rannveig Skrede

The present invention provides a synthetic mRNA leader for enhancing the expression of a gene encoding a desired protein, vectors comprising said synthetic mRNA leader and methods of producing a desired gene product using said synthetic mRNA leader and vector, said method comprising expressing said gene using a synthetic mRNA leader which comprises from 5' to 3&#x2032;: (i) a first mRNA leader sequence element; (ii) a spacer region; and (iii) a second mRNA leader sequence element; wherein said first mRNA leader sequence element is a modified transcription-stimulating mRNA leader capable of enhancing transcription of a gene relative to an unmodified reference mRNA leader sequence and/or said second mRNA leader element is a modified translation-stimulating mRNA leader capable of enhancing the translation of a gene transcript relative to an unmodified reference mRNA leader sequence.

Synthetic MRNA leaders

The present invention concerns a method of producing a desired gene product in a recombinant gene expression system, said method comprising expressing said gene from a Pm promoter-based expression system using at least two mutant elements selected from: (i) a mutant Pm promoter; (ii) a mutant mRNA leader; and (iii) a mutant XyIS; wherein said mutant elements each comprise one or more mutations which enhance expression of said desired gene. Particularly combinations of a mutant Pm promoter and a mutant mRNA leader are concerned. Isolated nucleic acid molecules, vectors, host cells, libraries, expression systems, methods of enhancing expression, obtaining nucleic acid molecules and identifying combination mutants which enhance expression, artificially constructed operons and their uses are also encompassed.

Enhanced expression from the Pm promoter

The present invention concerns a method of producing a desired heterologous gene product wherein said heterologous gene product is expressed from a strong promoter, said method comprising expressing said gene using a mutant mRNA leader which comprises one or more mutations which enhance transcription of the gene. The invention also provides a mutant Pm mRNA leader sequence, and a vector and a library comprising the leader sequence. Methods of obtaining an mRNA mutant leader and identifying an mRNA mutant leader are encompassed, along with a vector for selection or identification of an mRNA leader mutant and a use thereof for screening.

Enhanced expression method

The invention provides a nucleic acid molecule comprising: (a) a nucleotide sequence as shown in SEQ ID No. 35; or (b) a nucleotide sequence which is the complement of SEQ ID No. 35; or (c) a nucleotide sequence which is degenerate with SEQ ID No. 35; or (d) a nucleotide sequence hybridising under conditions of high stringency to SEQ ID No. 35, to the complement of SEQ ID No. 35, or to a hybridisation probe derived from SEQ ID No. 35 or the complement thereof; or (e) a nucleotide sequence having at least 80% sequence identity with SEQ ID No. 35; or (f) a nucleotide sequence having at least 65% sequence identity with SEQ ID No. 35 wherein said sequence preferably encodes or is complementary to a sequence encoding a nystatin PKS enzyme or a part thereof. Also provided are part of such molecules and polypeptides (and parts thereof) encoded by such a nucleic acid molecule, and the use of such molecules and polypeptides in facilitating nystatin biosynthesis and in the synthesis of nystatin derivatives and novel polyketide as macrolide structures.

Genes encoding a nystatin polyketide synthase and their manipulation and utility

It is described biologically pure bacterial cultures of mutant strains of, which produces large amounts of alginate. The alginate may contain a certain determined content of mannuronate and guluronate residues, possible presence and determined level of acetyl groups in the alginate, and a desired molecular weight of the alginate. Also high yielding mutants with regulation of alginate production, is described. The invention further provides methods for producing new mutant strains ofand variants thereof, and use the resulting strains in alginate production.

[object Object]

The invention provides a nucleic acid molecule comprising: (a) a nucleotide sequence as shown in SEQ ID No. 35; or (b) a nucleotide sequence which is the complement of SEQ ID No. 35; or (c) a nucleotide sequence which is degenerate with SEQ ID No. 35; or (d) a nucleotide sequence hybridising under conditions of high stringency to SEQ ID No. 35, to the complement of SEQ ID No. 35, or to a hybridisation probe derived from SEQ ID No. 35 or the complement thereof; or (e) a nucleotide sequence having at least 80% sequence identity with SEQ ID No. 35; or (f) a nucleotide sequence having at least 65% sequence identity with SEQ ID No. 35 wherein said sequence preferably encodes or is complementary to a sequence encoding a nystatin PKS enzyme or a part thereof. Also provided are part of such molecules and polypeptides (and parts thereof) encoded by such a nucleic add molecule, and the use of such molecules and polypeptides in facilitating nystatin biosynthesis and in the synthesis of nystatin derivatives and novel polyketide as macrolide structures.

The present invention provides an expression vector comprising an RK2 minimum replicon together with an expression cassette comprising the regulatory functions of a TOL plasmid, and, in particular, an expression vector comprising a RK2 minimum replicon together with a promoter Pm and/or Pu and a corresponding regulatory gene xylS and/or xylR as derived from a TOL plasmid. Such expression vectors may be used to express desired genes in a wide range of gram negative and gram positive bacterial hosts.

Expression vectors based on RK2 and TOL plasmids

Growing community of inventors

Trondheim, Norway

Svein Valla