A chromosomal expression vector for Escherichia coli based on the bacteriophage Mu.
A new Escherichia coli expression vector with increased stability was developed based on bacteriophage Mu. Unlike traditional expression vectors, the vector described herein is chromosome based rather than existing as an autonomously replicating plasmid. The chromosomal location resulted in extreme stability of the vector even in the absence of selective pressure. Both replication and heterologous protein synthesis could be induced by temperature shift. Expression of the heterologous gene was controlled by the Mu middle promoter and was dependent on the presence of the transactivator, Mor, of the Mu middle promoter. Four proteins, beta-galactosidase, chloramphenicol acetyltransferase, porcine somatotropin and human growth hormone, were made from this vector at levels ranging from 5 to 20% of total cell protein. Expression from the middle promoter was highest when inductions were done in rich media. The expression of some genes varied in different strains.
Weinberg
R A
RA
Monsanto Company, St. Louis, MO 63198.
De Ciechi
P A
PA
Obukowicz
M
M
D000818:Animals; D010583:Bacteriophage mu / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D003470:Culture Media; D004277:DNA, Single-Stranded; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D014644:Genetic Variation; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000037:antagonists & inhibitors; D013552:Swine; D013696:Temperature; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000037:antagonists & inhibitors; D054334:Viral Regulatory and Accessory Proteins
D003470:Culture Media; D004277:DNA, Single-Stranded; D012097:Repressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins