Process for production of polypeptides

Abstract

Vectors for producing polypeptides heterologous to prokaryotes are described comprising, along with the polypeptide-encoding nucleic acid, anti-termination nucleic acid that inhibits intragenic transcription termination with a non-lambda promoter therefor and/or nucleic acid encoding a GreA or GreB protein and a promoter therefor. Also described are processes for producing a heterologous polypeptide in prokaryotic host cells utilizing such elements to improve the quality and/or quantity of heterologous polypeptide produced.

1 . A vector comprising nucleic acid encoding GreA or GreB protein, nucleic acid encoding a polypeptide heterologous to prokaryotic cells, and one or more promoters for the nucleic acids.  
     
     
         2 . The vector of  claim 1  wherein the nucleic acid encodes GreB.  
     
     
         3 . The vector of  claim 1  wherein the prokaryotic cells are bacterial cells.  
     
     
         4 . The vector of  claim 1  wherein the polypeptide is a mammalian polypeptide.  
     
     
         5 . A process for producing a heterologous polypeptide in prokaryotic host cells comprising: 
 (a) culturing the host cells, which comprise nucleic acid encoding GreA or GreB protein, nucleic acid encoding the heterologous polypeptide, and one or more promoters for the nucleic acids; and    (b) recovering the heterologous polypeptide from the cells or from cell culture medium.    
     
     
         6 . The process of  claim 5  wherein nucleic acid encoding GreB protein is expressed.  
     
     
         7 . The process of  claim 5  wherein the cells are bacterial cells.  
     
     
         8 . The process of  claim 5  wherein the heterologous polypeptide is a mammalian polypeptide.  
     
     
         9 . The process of  claim 5  wherein the mammalian polypeptide is a human polypeptide.  
     
     
         10 . The process of  claim 9  wherein the human polypeptide is thrombopoietin (TPO) or fibroblast growth factor-5 (FGF-5).  
     
     
         11 . The process of  claim 5  wherein the promoter is a trp or alkaline phosphatase promoter or both.  
     
     
         12 . The process of  claim 5  wherein the polypeptide is recovered from the cytoplasm or periplasm of the cells.  
     
     
         13 . The process of  claim 5  wherein the polypeptide is recovered from the cell culture medium.