Concatemers of differentially expressed multiple genes

Details Concatemers of differentially expressed multiple genes Concatemers of differentially expressed multiple genes

C - Chemistry, Metallurgy

12

N

C12N 15/66 (2006.01) C12N 15/10 (2006.01) C12N 15/70 (2006.01) C12N 15/81 (2006.01) C12N 15/85 (2006.01)

Patent

CA 2474161

In the present invention are disclosed concatemers of concatenated expression cassettes and vectors that enable the synthesis of such concatemers. The concatemer comprises in the 5' .fwdarw.3' direction a cassette of nucleotide sequence of the general formula [rs2-SP-PR-X-TR-SP-rs1]n wherein rs1 and rs2 together denote a functional restriction site, SP individually denotes a spacer of at least two nucleotide bases, PR denotes a promoter, capable of functioning in a cell, X denotes an expressible nucleotide sequence, TR denotes a terminator, and SP individually denotes a spacer of at least two nucleotide bases, and n >= 2, and wherein at least a first cassette is different from a second cassette. The main purpose of these concatemers is the controllable and co-ordinated expression of large numbers of heterologous genes in a single host. Furthermore, the invention relates to a concatemer of cassettes of nucleotide sequences and a method for preparing the concatemers. In a further aspect, the invention relates to transgenic host cells comprising at least one concatemer according to the invention, as well as to a method for preparing the transgenic host cells. Finally, the invention relates to a vector comprising a cassette of nucleotides, a method for preparing said vector, a nucleotide library comprising at least two primary vectors each comprising a cassette of nucleotides, a method for preparing the library.

L'invention concerne des concatémères de cassettes d'expression concaténées et des vecteurs qui permettent la synthèse de tels concatémères. Le but principal de ces concatémères est l'expression maîtrisable et coordonnée de grands nombres de gènes hétérologues dans un hôte unique. L'invention concerne également un concatémère de cassettes de séquences de nucléotides, ainsi qu'un procédé de préparation de ces concatémères. Dans un autre aspect, l'invention concerne des cellules hôtes transgéniques comprenant au moins un concatémère de cette invention, ainsi qu'un procédé de préparation de ces cellules hôtes transgéniques. Enfin, l'invention concerne un vecteur comprenant une cassette de nucléotides, un procédé de préparation de ce vecteur, une banque de nucléotides comprenant au moins deux vecteurs primaires comprenant chacun une cassette de nucléotides, et un procédé de préparation de cette banque.

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