Abstract: The present invention provides DNA compounds that encode the expandase/hydroxylase enzyme of Cephalosporium acremonium. The compounds can be used to construct recombinant DNA expression vectors for a variety of host cells, including E. coli, Penicillium, and Cephalosporium.

Abstract: The invention provides isolated nucleic acid compounds encoding a multiple drug resistance protein of Aspergillus fumigatus. Vectors and transformed host cells comprising the multiple drug resistance-encoding DNA of Aspergillus fumigatus AfuMDR1 are also provided. The invention further provides assays which utilize these transformed host cells.

Abstract: The invention provides isolated nucleic acid compounds encoding a multiple drug resistance protein of Cryptococcus neoformans. Vectors and transformed host cells comprising the multiple drug resistance-encoding DNA of Cryptococcus neoformans CneMDR1 are also provided. The invention further provides assays which utilize these transformed host cells.

Abstract: A method for increasing the antibiotic-producing ability of an antibiotic-producing microbial host cell is disclosed. The method involves transforming an antibiotic-producing microorganism with a recombinant DNA cloning vector that codes for the expression of an antibiotic biosynthetic enzyme or other gene product. The gene preferably codes for a product that is rate-limiting in the antibiotic biosynthetic pathway in Streptomyces. Plasmids that are useful for increasing tylosin production in accordance with the present method include plasmids pHJL280, pHJL284, pHJL309, pHJL311, and pHJL315. The invention further comprises microorganisms transformed with plasmids pHJL280, pHJL284, pHJL309, pHJL311, and pHJL315 and also other microorganisms and vectors used in accordance with the present method.

Abstract: The instant invention provides the femA gene of Staphylococcus epidermidis and all degenerate sequences thereof, the protein encoded by the femA gene (FemA), and vectors and microorganisms comprising genes encoding the FemA protein.

Abstract: The present invention provides a method for transforming an actinomycete with an integrating vector which has the advantages of high transformation rates into a broad host range, site-specific integration, and stable maintenance without antibiotic selection. Also provided are methods for the increased production of antibiotics and for the production of hybrid antibiotics.

Abstract: Provided are gene sequences encoding tylosin biosynthetic gene products. In particular, recombinant DNA vectors comprising DNA sequences encoding the tylA, tylB, tylI and tylG activities of Streptomyces fradiae are provided. Also provided are host cells transformed with the noted vectors and a method for increasing the tylosin-producing ability of a tylosin-producing organism.

Abstract: Spiramycin antibiotic biosynthetic genes of Streptomyces ambofaciens are provided by the present invention, in addition to a variety of recombinant DNA vectors. The genes also function in other macrolide producing organisms. The genes can be used to increase or otherwise alter the macrolide antibiotic-producing ability of an organism. The present invention also provides host strains comprising mutant spiramycin biosynthetic genes which can be used to generate novel antibiotics. Also provided is a method for preparing the mutant gene comprising mutating cloned spiramycin biosynthetic DNA by transposon mutagenesis with subsequent transformation into a macrolide-antibiotic producing host and homologous recombination into its genome, to generate stable mutant cell lines.

Abstract: The present invention provides DNA compounds that encode deacetoxycephalosporin C synthetase (DAOCS) activity. The compounds can be used to construct recombinant DNA expression vectors for a wide variety of host cells, including E. coli, Penicillium, Streptomyces, Aspergillus, and Cephalosporium.

Abstract: The recovery of vitamin K-dependent proteins produced by transformed microorganisms can be effected from the cell culture medium utilizing the changes in the protein which occur in the presence of divalent cations. The present process uses divalent cations to alter the binding affinity of the proteins and thereby selectively elute the proteins away from contaminants in the culture medium using standard chromatography.

Abstract: DNA compounds encoding the Streptomyces lipmanii isopenicillin N synthetase (IPNS) gene are useful for constructing a variety of recombinant DNA vectors. The vectors are useful in producing IPNS in a wide variety of host cells, such as Streptomyces, Penicillium, and Cephalosporium. DNA compounds encoding the transcription and translation activating sequence and transcription termination sequence of the S. lipmanii IPNS gene are also useful in the construction of expression vectors, especially Streptomyces expression vectors. The S. lipmanii IPNS gene can be isolated from plasmid pOGO239, available from the Northern Regional Research Center under accession number NRRL B-18250.

Abstract: The present invention is a method for isolating antibiotic biosynthetic genes. To practice the method, an antibiotic resistance-conferring DNA segment is labelled and used as a probe to find, via DNA hydridization, homologous DNA in a genetic library which comprises chromosomal and plasmid DNA of an antibiotic-producing organism. Individual vectors of the genetic library which hybridize to the antibiotic resistance-conferring gene, and which comprise .about.1-45 kb of contiguous DNA from the antibiotic-producing organism, which also comprise an antibiotic biosynthetic gene. The present method is exemplified by using the erythromycin resistance-conferring gene of Streptomyces erythreus to clone the erythromycin biosynthetic pathway from the same organism. The erythromycin biosynthetic pathway isolated with the present method synthesizes erythromycin when introduced into S. lividans TK23.