Plasmid PXL3179 or NV1FGF

Abstract

A prokaryotic recombinant host cell comprising a heterologous replication initiation protein that activates a conditional origin of replication and an extrachromosomal DNA molecule comprising a heterologous therapeutic gene and a conditional origin of replication whose functionality in the prokaryotic recombinant host cell requires a replication initiating protein which is foreign to the host cell is described. The host cell may comprise a pir gene having at least one mutation, which may occur in the pir gene copy number control region, the pir gene leucine zipper-like motif, or the pir gene DNA binding region.

1 - 49 . (canceled) 
     
     
         50 . Plasmid pXL3179. 
     
     
         51 . A composition comprising the plasmid of  claim 50  and a vehicle. 
     
     
         52 . A pharmaceutical composition comprising the plasmid of  claim 50  and a pharmaceutically acceptable vehicle. 
     
     
         53 . A recombinant host cell comprising the plasmid of  claim 50 . 
     
     
         54 . The recombinant host cell of  claim 53 , wherein the host cell expresses a n initiator protein. 
     
     
         55 . The recombinant host cell of  claim 54 , wherein the host cell is  Escherichia coli.    
     
     
         56 . The recombinant host cell of  claim 54 , wherein the host cell is TEX1, as deposited under Collection Nationale de Cultures de Microorganismes accession number CNCM 1-3569. 
     
     
         57 . A method of producing plasmid pXL3179, comprising
 a) culturing a recombinant host cell according to  claim 53  under conditions permitting replication of the plasmid, and   b) isolating the plasmid produced by the host cell.   
     
     
         58 . A plasmid comprising the nucleotide sequence set forth in SEQ ID NO: 40. 
     
     
         59 . A composition comprising the plasmid of  claim 58  and a vehicle. 
     
     
         60 . A pharmaceutical composition comprising the plasmid of  claim 58  and a pharmaceutically acceptable vehicle. 
     
     
         61 . A recombinant host cell comprising the plasmid of  claim 58 . 
     
     
         62 . The recombinant host cell of  claim 61 , wherein the host cell expresses a π initiator protein. 
     
     
         63 . The recombinant host cell of  claim 62 , wherein the host cell is  Escherichia coli.    
     
     
         64 . The recombinant host cell of  claim 62 , wherein the host cell is TEX1, as deposited under Collection Nationale de Cultures de Microorganismes accession number CNCM I-3569. 
     
     
         65 . A method of producing a plasmid comprising the nucleotide sequence set forth in SEQ ID NO: 40, comprising
 a) culturing a recombinant host cell according to  claim 61  under conditions permitting replication of the plasmid, and   b) isolating the plasmid produced by the host cell.   
     
     
         66 . A plasmid consisting of the nucleotide sequence set forth in SEQ ID NO: 40. 
     
     
         67 . A composition comprising the plasmid of  claim 66  and a vehicle. 
     
     
         68 . A pharmaceutical composition comprising the plasmid of  claim 66  and a pharmaceutically acceptable vehicle. 
     
     
         69 . A recombinant host cell comprising the plasmid of  claim 66 . 
     
     
         70 . The recombinant host cell of  claim 69 , wherein the host cell expresses a T initiator protein. 
     
     
         71 . The recombinant host cell of  claim 70 , wherein the host cell is  Escherichia coli.    
     
     
         72 . The recombinant host cell of  claim 70 , wherein the host cell is TEX1, as deposited under Collection Nationale de Cultures de Microorganismes accession number CNCM 1-3569. 
     
     
         73 . A method of producing a plasmid consisting of the nucleotide sequence set forth in SEQ ID NO: 40, comprising
 a) culturing a recombinant host cell according to  claim 69  under conditions permitting replication of the plasmid, and   b) isolating the plasmid produced by the host cell.