BIOMAT Database Logo BIOMAT Database Logo

Improved expression of a hybrid Streptomyces clavuligerus cefE gene in Penicillium chrysogenum. 1994

S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285.

A hybrid cefE gene, encoding penicillin N expandase, was constructed by fusing the promoter sequences, Pcp, and terminator sequences, Pct from the Penicillium chrysogenum pcbC gene to the open reading frame (orf), cefEorf, from the Streptomyces clavuligerus cefE gene. The resulting hybrid gene, Pcp/cefE'orf/Pct, differed from a previously reported hybrid cefE gene contained on plasmid pPS65. The latter gene, Pcp/cefE'orf/Sct, contained the Pcp sequences fused to the S. clavuligerus cefE orf still attached to the S. clavuligerus terminator sequences, Sct. The new hybrid gene was transformed into P. chrysogenum on plasmid vector pRH6. Transformants were selected by phleomycin resistance conferred by a hybrid ble gene present on plasmid pRH6. The hybrid ble gene was formed by attaching Pcp sequences to the ble orf. Among transformants obtained with pRH6, one exhibited a 70-fold higher level of activity of penicillin N expandase than the best transformant previously obtained from a 10-fold larger population of pPS65 transformants. The penicillin N expandase activity in pRH6 transformant, 9EN-5-1, was fourfold higher than the activity in the S. clavuligerus strain used as the source of the cefE orf and 75% of the activity observed in an industrial strain of Cephalosporium acremonium. Sequencing of the junctions of the heterologous DNA in Pcp/cefEorf/Pct uncovered a modification of the cefE open reading frame introduced during construction of the hybrid gene; the modified open reading frame is designated cefE'orf.

UI MeSH Term Description Entries
D007535 Isomerases A class of enzymes that catalyze geometric or structural changes within a molecule to form a single product. The reactions do not involve a net change in the concentrations of compounds other than the substrate and the product.(from Dorland, 28th ed) EC 5. Isomerase
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D010407 Penicillium A mitosporic Trichocomaceae fungal genus that develops fruiting organs resembling a broom. When identified, teleomorphs include EUPENICILLIUM and TALAROMYCES. Several species (but especially PENICILLIUM CHRYSOGENUM) are sources of the antibiotic penicillin. Penicilliums
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D002511 Cephalosporins A group of broad-spectrum antibiotics first isolated from the Mediterranean fungus ACREMONIUM. They contain the beta-lactam moiety thia-azabicyclo-octenecarboxylic acid also called 7-aminocephalosporanic acid. Antibiotics, Cephalosporin,Cephalosporanic Acid,Cephalosporin,Cephalosporin Antibiotic,Cephalosporanic Acids,Acid, Cephalosporanic,Acids, Cephalosporanic,Antibiotic, Cephalosporin,Cephalosporin Antibiotics
D004274 DNA, Recombinant Biologically active DNA which has been formed by the in vitro joining of segments of DNA from different sources. It includes the recombination joint or edge of a heteroduplex region where two recombining DNA molecules are connected. Genes, Spliced,Recombinant DNA,Spliced Gene,Recombinant DNA Research,Recombination Joint,DNA Research, Recombinant,Gene, Spliced,Joint, Recombination,Research, Recombinant DNA,Spliced Genes
D005798 Genes, Bacterial The functional hereditary units of BACTERIA. Bacterial Gene,Bacterial Genes,Gene, Bacterial
D005818 Genetic Engineering Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc. Genetic Intervention,Engineering, Genetic,Intervention, Genetic,Genetic Interventions,Interventions, Genetic
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA

Related Publications

S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus: Growth yields and morphological characterization.
August 2003, Biotechnology and bioengineering,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Characteristics of the beta-lactam synthesizing enzymes of Streptomyces clavuligerus, Cephalosporium acremonium, and Penicillium chrysogenum.
January 1988, Annals of the New York Academy of Sciences,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Characterization and expression in Streptomyces lividans of cefD and cefE genes from Nocardia lactamdurans: the organization of the cephamycin gene cluster differs from that in Streptomyces clavuligerus.
January 1993, Molecular & general genetics : MGG,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus: Kinetics of adipoyl-7-aminodeacetoxycephalosporanic acid and byproduct formations.
August 2003, Biotechnology and bioengineering,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Physiological characterisation of Penicillium chrysogenum strains expressing the expandase gene from Streptomyces clavuligerus during batch cultivations. Growth and adipoyl-7-aminodeacetoxycephalosporanic acid production.
October 2001, Applied microbiology and biotechnology,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Regulated system for heterologous gene expression in Penicillium chrysogenum.
February 1997, Applied and environmental microbiology,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Expression of the trpC gene from Penicillium chrysogenum in Aspergillus nidulans.
October 1989, Biotechnology and applied biochemistry,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Influence of the adipate and dissolved oxygen concentrations on the beta-lactam production during continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus.
January 2003, Metabolic engineering,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
[Penicillium chrysogenum genetics. I. Heterokaryosis in Penicillium chrysogenum].
January 1954, Rendiconti - Istituto superiore di sanita,
S W Queener, and R J Beckmann, and C A Cantwell, and R L Hodges, and D L Fisher, and J E Dotzlaf, and W K Yeh, and D McGilvray, and M Greaney, and P Rosteck
Glycerol utilization gene cluster in Streptomyces clavuligerus.
May 2009, Applied and environmental microbiology,
Copied contents to your clipboard!