PCR-cloning of the pac gene from E. coli ATCC11105--observations on the expression of a recombinant periplasmic protein

The penicillin acylase gene (pac) amplified by polymerase chain reaction (PCR) from Escherichia coli (E.coli) ATCC11105 genomic DNA was cloned into pUC9, pEMBL9+ and pCP40 vectors. A penicillin acylase precursor was overexpressed at 26 degrees C from pac-pEMBL9+ and pac-pCP40 constructs transformed...

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Veröffentlicht in:	Biochemistry and molecular biology international 1996-02, Vol.38 (2), p.315-324
Hauptverfasser:	Gümüşel, F, Kiremit, N, Işsever, S, Oğras, T T, Bermek, E
Format:	Artikel
Sprache:	eng
Schlagworte:	Base Sequence Cloning, Molecular Enzyme Induction Escherichia coli - enzymology Escherichia coli - genetics Gene Expression Regulation, Bacterial Genes, Bacterial - genetics Molecular Sequence Data Molecular Weight Penicillin Amidase - biosynthesis Penicillin Amidase - chemistry Penicillin Amidase - genetics Polymerase Chain Reaction - methods Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - chemistry Temperature
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Zusammenfassung:	The penicillin acylase gene (pac) amplified by polymerase chain reaction (PCR) from Escherichia coli (E.coli) ATCC11105 genomic DNA was cloned into pUC9, pEMBL9+ and pCP40 vectors. A penicillin acylase precursor was overexpressed at 26 degrees C from pac-pEMBL9+ and pac-pCP40 constructs transformed into E.coli strains JM109 and pCI857 containing HB101, respectively. With the thermo-inducible pac-pCP40 construct some level of mature alpha and beta subunits and varying degrees of enzyme activity were also detected.
ISSN:	1039-9712