TY - JOUR
T1 - Cloning and nucleotide sequence determination of the isopenicillin N synthetase gene from Streptomyces clavuligerus
AU - Leskiw, Brenda K.
AU - Aharonowitz, Yair
AU - Mevarech, Moshe
AU - Wolfe, Saul
AU - Vining, Leo C.
AU - Westlake, Donald W.S.
AU - Jensen, Susan E.
N1 - Funding Information:
This work has been supported by the Natural Sciences and Engineering Research Council of Canada, and by the Alberta Heritage Foundation For Medical Research. We thank Ms. Pamela Ban-ser and Ms. Annie Wong for technical assistance, and Dr. James L. Doran for helpful discussions during the preparation of this manuscript.
PY - 1988/2/29
Y1 - 1988/2/29
N2 - The isopenicillin N synthetase (IPNS) gene from Streptomyces clavuligerus was isolated from an Escherichia coli plasmid library of S. clavuligerus genomic DNA fragments using a 44-mer mixed oligodeoxynucleotide probe. The nucleotide sequence of a 3-kb region of the cloned fragment from the plasmid, pBLl, was determined and analysis of the sequence showed an open reading frame that could encode a protein of 329 amino acids with an Mr of 36 917. When the S. clavuligerus DNA from pBLl was introduced into an IPNS-deficient mutant of S. clavuligerus on the Streptomyces vector pIJ941, the recombinant plasmid was able to complement the mutation and restore IPNS activity. The protein coding region of the S. clavuligerus IPNS gene shows about 63% and 62% similarity to the Cephalosporium acremonium and Penicillium chrysogenum IPNS nucleotide sequences, respectively, and the predicted amino acid sequence of the encoded protein showed about 56% similarity to both fungal sequences.
AB - The isopenicillin N synthetase (IPNS) gene from Streptomyces clavuligerus was isolated from an Escherichia coli plasmid library of S. clavuligerus genomic DNA fragments using a 44-mer mixed oligodeoxynucleotide probe. The nucleotide sequence of a 3-kb region of the cloned fragment from the plasmid, pBLl, was determined and analysis of the sequence showed an open reading frame that could encode a protein of 329 amino acids with an Mr of 36 917. When the S. clavuligerus DNA from pBLl was introduced into an IPNS-deficient mutant of S. clavuligerus on the Streptomyces vector pIJ941, the recombinant plasmid was able to complement the mutation and restore IPNS activity. The protein coding region of the S. clavuligerus IPNS gene shows about 63% and 62% similarity to the Cephalosporium acremonium and Penicillium chrysogenum IPNS nucleotide sequences, respectively, and the predicted amino acid sequence of the encoded protein showed about 56% similarity to both fungal sequences.
KW - Cephalosporium acremonium
KW - Penicillium chrysogenum
KW - Recombinant DNA
KW - mutant complementation
KW - pUC119 library
KW - β-lactam biosynthesis
UR - http://www.scopus.com/inward/record.url?scp=0023848579&partnerID=8YFLogxK
U2 - 10.1016/0378-1119(88)90557-4
DO - 10.1016/0378-1119(88)90557-4
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
AN - SCOPUS:0023848579
SN - 0378-1119
VL - 62
SP - 187
EP - 196
JO - Gene
JF - Gene
IS - 2
ER -