Enhanced Expression Method
AssigneeLeif Eiriksson Nyskaping AS Corporation
InventorsSvein Valla, Rahmi Lale, Laila Berg, Ingrid Bakke
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Abstract
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.
Claims34
A method of producing a desired heterologous gene product wherein a gene encoding 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 said gene.
A Pm mRNA leader sequence which comprises one or more mutations as compared to the wildtype Pm mRNA leader sequence wherein said mutant mRNA leader sequence is capable of enhancing expression of a desired gene product.
A method of obtaining a mRNA mutant leader capable of enhancing the transcription of a desired gene, said method comprising introducing one or more mutations into the DNA corresponding to an mRNA leader, expressing said desired gene using said mRNA mutant leader in a host cell and selecting an mRNA leader mutant which enhances transcription of the said desired gene.
A method of identifying an mRNA leader mutant which enhances expression of a desired gene, said method comprising:
- providing a vector comprising a promoter, a said desired gene under the control of said promoter and a reporter gene translationally coupled thereto;
- introducing a DNA sequence corresponding to said mRNA leader mutant into said vector upstream of said desired gene;
- introducing said vector into a host cell and determining the level of expression of said reporter gene.
A vector for selection or identification of an mRNA leader mutant which comprises one or more mutations which enhance transcription of a gene, said vector comprising a promoter, a desired gene under the control of said promoter and a reporter gene translationally coupled thereto and a site for insertion of a DNA region corresponding to the said leader mutant upstream of said desired gene.
Dependencies are inferred from the claim text.