Enhanced expression from the Pm promoter

摘要

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.

主权项

1. A method of producing a desired gene product in a recombinant gene expression system wherein transcript production from said gene is increased and results in enhanced expression of the desired gene product, said method comprising expressing said gene with a Pm promoter-based expression system using:
(i) a mutant Pm promoter which increases the amount of transcript produced and enhances gene expression compared to gene expression obtained using the corresponding unmutated Pm promoter, wherein said mutant Pm promoter comprises a sequence selected from the group consisting of SEQ ID NOS: 102 to 119, 121 to 137 and 144 to 145, and (ii) a mutant Pm mRNA leader which increases the amount of transcript produced and enhances gene expression compared to gene expression obtained using the corresponding unmutated Pm mRNA leader, wherein said mutant Pm mRNA leader comprises a sequence selected from the group consisting of SEQ ID NOS: 2 to 12, 14 to 37 and 147 to 163, wherein said mutant Pm promoter and mutant Pm mRNA leader enhance gene expression compared to gene expression obtained using only the mutant Pm promoter and compared to gene expression obtained using only the mutant Pm mRNA leader.