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A series of new plasmid expression vectors (the pTrc series) has been constructed for the regulated expression of genes in Escherichia coli. Based on pKK233-2 [Amann and Brosius, Gene 40 (1985) 183-190], the vectors carry a strong hybrid trp/lac promoter, the lacZ ribosome-binding site (RBS), the multiple cloning site of pUC18 and the rrnB transcription terminators. With the aid of synthetic oligodeoxynucleotides, the multiple cloning site has been inserted behind an NcoI site in three reading frames. Thus, the vectors are equally useful for the expression of proteins in their authentic, non-fused form (by using the NcoI site) and for the expression of fusion proteins (by choosing any of the cloning sites in the correct translational frame). To ensure complete repression of the hybrid trp/lac promoter during construction and growth in any host strain, the lacIq allele of the lac repressor gene was added to some of the vectors. The complete vector nucleotide sequence and examples of heterologous gene expression (human coagulation factor XIIIa and human placental anticoagulant protein PP4) with the new vectors are presented.
E Amann, and
B Ochs, and
K J Abel
January 1986,
Gene,
E Amann, and
B Ochs, and
K J Abel
November 1983,
Gene,
E Amann, and
B Ochs, and
K J Abel
April 1988,
Gene,
E Amann, and
B Ochs, and
K J Abel
May 1996,
Analytical biochemistry,
E Amann, and
B Ochs, and
K J Abel
October 1989,
BioTechniques,
E Amann, and
B Ochs, and
K J Abel
January 1987,
Gene,
E Amann, and
B Ochs, and
K J Abel
April 2010,
Current microbiology,
E Amann, and
B Ochs, and
K J Abel
August 2010,
Applied and environmental microbiology,
E Amann, and
B Ochs, and
K J Abel
March 2006,
BioTechniques,
E Amann, and
B Ochs, and
K J Abel
January 2011,
Journal of biotechnology,
