Expression vectors with modified ColE1 origin of replication for control of plasmid copy number
AssigneeBoehringer Ingelheim International GmbH
InventorsKarl Bayer, Reingard Grabherr, Erik Nilsson, Gerald Striedner
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Abstract
In an expression vector having a ColE1 replication system, the homology of the RNAI and RNAII of the ColE1 origin of replication to uncharged tRNAs is modified mutations in the coding region of the RNAI gene and corresponding mutations in the RNAII gene. The mutation results in one or more base exchanges in loop 1 and/or loop 2 and/or loop 3 of RNAI and RNAII. In methods using this vector for producing recombinant proteins, plasmid copy number is stably maintained. In methods for plasmid production, high plasmid copy numbers can be obtained.
Claims14
An expression vector having a Col E1 replication system, wherein the homology of the RNAI and RNAII of the ColE1 origin of replication to one or more uncharged tRNAs is modified by one or more mutations in the coding region of the RNAI gene and one or more corresponding mutations in the RNATI gene, said mutation(s) resulting in one or more base exchanges in loop 1 and/or loop 2 and/or loop 3 of RNAI and RNAII.
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