An Escherichia coli expression vector for high-level production of heterologous proteins in fusion with CMP-KDO synthetase.
The construction of a vector which overproduces the enzyme, CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyl-transferase (CMP-KDO synthetase or CKS) and its use as an expression vector for producing heterologous proteins in E. coli is described. The vector, which includes a modified lac promoter and synthetic ribosome binding site upstream of the native kdsB gene (encoding CKS), produces CKS at levels as high as 70% of the total cellular proteins. Several heterologous gene sequences have been fused to the 3'-end of the kdsB gene with resulting protein fusions produced at a level of up to 40% of the total cellular proteins.
Bolling
T J
TJ
Corporate Molecular Biology, Abbott Laboratories, Abbott Park, IL 60064.
Mandecki
W
W
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins
D015700:HIV Envelope Protein gp41; D011993:Recombinant Fusion Proteins; D009713:Nucleotidyltransferases; C028984:3-deoxy-manno-octulosonate cytidyltransferase