Abbott Laboratories Corporation

University of British Columbia

Baylor College of Medicine

Leonard Katz

James B McAlpine

Richard G Summers, Jr

Diane L Stassi

Ana Pereda-Lopez

Stefano Donadio

Stephan J Kakavas

Xiaoan Ruan

James Tuan

James R Lupski

Julian E Davies

Leland Shawn Paul

Stephen B Delcardayre

Michael J Staver

Charles Lester Ginsburgh

Lisa Beth Cohen

Annapur Gurulingappa Shivakumar

Rita Irene Vanags

The invention provides novel erythromycin derivatives in which methyl groups on the macrolactone ring have been substituted with &mdash;H, -Et, and/or &mdash;OH. The invention also provides reagents such as isolated polynucleotides, vectors comprising the polynucleotides and host cells transformed with the vectors for making the novel compounds. Methods for making the compounds utilizing genetic engineering techniques are also disclosed.

Polyketide derivatives and recombinant methods for making same

An isolated and purified Coenzyme A disulfide reductase (CoADR) is provided. Oligonucleotides encoding the CoADR, vectors and host cells containing such oligonucleotides are also provided. In addition, antibodies reactive with the CoADR are provided, as are methods of isolating the CoADR, producing recombinant CoADR, using CoADR for screening compounds for CoADR-modulating activity, and detecting organisms which produce CoADR a test sample. Methods for identifying a gene encoding a CoADR are also provided.

Coenzyme A disulfide reductase, and inhibitors thereof useful as

The invention provides novel erythromycin derivatives in which methyl groups on the macrolactone ring have been substituted with --H, -Et, and/or --OH and the ethyl side-chain has been substituted with hydroxymethyl or dihydroxycyclohexylmethyl side-chains. The invention also provides reagents such as isolated polynucleotides, vectors comprising the polynucleotides and host cells transformed with the vectors for making the novel compounds. Methods for making the compounds utilizing genetic engineering techniques are also disclosed.

The invention provides novel erythromycin derivatives in which methyl groups on the macrolactone ring have been substituted with --H, --Et, and/or --OH. The invention also provides reagents such as isolated polynucleotides, vectors comprising the polynucleotides and host cells transformed with the vectors for making the novel compounds. Methods for making the compounds utilizing genetic engineering techniques are also disclosed.

A method to produce novel polyketide structures by designing and introducing specified changes in the DNA governing the synthesis of the polyketide is disclosed. The biosynthesis of specific polyketide analogs is accomplished by genetic manipulation of a polyketide-producing microorganism by isolating a polyketide biosynthetic gene-containing DNA sequence, identifying enzymatic activities associated within the DNA sequence, introducing one or more specified changes into the DNA sequence which codes for one of the enzymatic activities which results in an altered DNA sequence, introducing the altered DNA sequence into the polyketide-producing microorganism to replace the original sequence, growing a culture of the altered microorganism under conditions suitable for the formation of the specific polyketide analog, and isolating the specific polyketide analog from the culture. The method is most useful when the segment of the chromosome modified is involved in an enzymatic activity associated with polyketide biosynthesis, particularly for manipulating polyketide synthase genes from Saccarharopolyspora or Streptomyces.

Method of directing biosynthesis of specific polyketides

The present invention provides isolated polynucleotides from Saccharomyces erythraea that encode enzymes involved in the biosynthesis of polyketide-associated sugars. Methods of using the polynucleotides to produce novel glycosylation modified polyketides are also provided.

Polyketide-associated sugar biosynthesis genes

Recombinant DNA method for producing erythromycin analogs

Proteins heterologous to B. megaterium can be expressed in B. megaterium using a sporulation promoter from B. subtilis.

Expression of heterologous proteins in Bacillus megaterium utilizing

A method of interrupting the expression of a macromolecular synthesis operon in bacteria comprising the step of binding an antisense oligonucleotide to a single stranded DNA or to a mRNA transcribed from the macromolecular synthesis operon. The antisense oligonucleotide can be either sequence specific to a unique intergenic sequence or a sequence specific to a bacterial homologous sequence. By interrupting the expression of the macromolecular synthesis operon bacterial infections can be treated. Specific antisense oligonucleotides are disclosed. The ability of the antisense oligonucleotide to bind the mRNA or single stranded DNA also allows the identification of the bacteria by using a unique intergenic antisense oligonucleotide to bind to the single stranded DNA or to the mRNA transcribed from the macromolecular synthesis operon. A method for competitively inhibiting the protein products of the MMS operon with oligonucleotides is also disclosed. Methods of identifying unique intergenic sequence is also disclosed.

Antisense oligonucleotide antibiotics complementary to the

DNA cloning shuttle vectors, including a cosmid shuttle vector, for E. coli and Streptomyces are disclosed. Specifically, disclosed shuttle vectors pAL7002 (NRRL B-18055) and pNJ1 (NRRL B-18054) contain an E. coli origin of replication, Streptomyces replication functions, and antibiotic resistance markers for both E. coli and Streptomyces. In addition, pNJ1 contains a cos sequence. Novel 2-norerythromycin antibiotics A, B, C, and D, which were produced in a strain Streptomyces erythreus 12693-240 (NRRL B-18053) transformed by pNJ1 bearing DNA from Streptomyces antibioticus, are also disclosed. The present invention also provides a method for producing novel antibiotics. This method for antibiotic production is applied to the transformation of a blocked mutant of S. erythreus with genomic DNA from S. antibioticus but may be more broadly applied to genes of antibiotic-producing strains transformed into cells which are blocked in the pathway for production of a different antibiotic.

Biologically pure cultures of streptomyces and use thereof in macrolide

DNA cloning shuttle vectors, including a cosmid shuttle vector, for E. coli and Streptomyces are disclosed. Specifically, disclosed shuttle vectors pAL7002 (NRRL B-18055) and pNJl (NRRL B-18054) contain an E. coli origin of replication, Streptomyces replication functions, and antibiotic resistance markers for both E. coli and Streptomyces. In addition, pNJl contains a cos sequence. Novel 2-norerythromycin antibiotics A, B, C, and D, which were produced in a strain Streptomyces erythreus 12693-240 (NRRL B-18053) transformed by pNJl bearing DNA from Streptomyces antibioticus, are also disclosed. The present invention also provides a method for producing novel antibiotics. This method for antibiotic production is applied to the transformation of a blocked mutant of S. erythreus with genomic DNA from S. antibioticus but may be more broadly applied to genes to antibiotic-producing strains transformed into cells which are blocked in the pathway for production of a different antibiotic.

Growing community of inventors

Wheeling, IL, United States of America

Leonard Katz