Eli Lilly and Company

Ramachandra N Rao

Jan R Turner

Paul R Rosteck, Jr

Steven Kovacevic

Mark Alan Richardson

Stuart A Kuhstoss

Robert F Santerre

Stanley Gene Burgett

Keith A Otto

A DNA molecule isolated from Streptomyces ambofaciens encodes the multi-functional proteins which direct the synthesis of the polyketide platenolide.

Platenolide synthase gene

The present invention provides novel fusion proteins comprising cyclins and CDKs. A preferred embodiment of the invention provides fusion proteins comprising human cyclin D1 and human CDK4. The fusion proteins of the invention optionally contain modifications, which facilitate their purification. Addition of histidine residues to selected constructs allows purification via immobilized metal affinity chromatography. Antigenic determinants allowing monoclonal antibody-based affinity chromatography purification are provided in selected embodiments of the invention. Protease cleavage sites are incorporated in selected constructs to allow cleavage of the regions incorporated in the cyclin-CDK fusion proteins for purification. Additional modifications which facilitate purification include strepavadin binding domains and antigenic determinants for antibody affinity chromatography.

Fusion proteins comprising cell cycle regulatory proteins

A gene encoding activator protein (srmR) for increasing transcriptional efficiency of macrolide biosynthetic genes is disclosed and claimed. Methods for using srmR to increase macrolide biosynthetic gene transcription and identifying further macrolide biosynthetic pathways are disclosed. Recombinant DNA vectors comprising the srmR gene are disclosed.

Activator gene for macrolide biosynthesis

The present invention comprises novel recombinant DNA cloning and expression vectors which confer hygromycin B and/or G418 resistance to eukaryotic and prokaryotic host cells. The novel recombinant DNA vectors are derived from plasmid pKC203, a plasmid which can be isolated from E. coli JR225 (ATCC 31912). The hygromycin B and G418 resistance-conferring genes can be isolated on the 7.5 kb BglII restriction fragment or the 2.5 kb SalI-BglII restriction fragment of plasmid pKC203. The eukaryotic recombinant DNA vectors of the present invention are prepared by inserting such resistance-conferring restriction fragments into a vector, such as plasmid pSV5gpt, that comprises a eukaryotic promoter and the necessary functions for maintenance of the vector as an extrachromosomal element or as an integrated sequence in the host cell chromosomal DNA. Furthermore, the present invention comprises useful derivatives of plasmid pKC203 which, although comprising no eukaryotic elements, are useful recombinant vectors for E. coli and other prokaryotes and serve as starting material for the construction of eukaryotic vectors that confer hygromycin B and/or G418 resistance to eukaryotic host cells. One useful derivative of plasmid pKC203 is constructed by circularizing the .about.7.5 kb BglIII restriction fragment of plasmid pKC203 to form plasmid pSC701, which can be further digested with HaeII or SauIIIA1 to form smaller plasmids. The present invention also comprises the novel transformants of the aforementioned recombinant DNA vectors.

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Indianapolis, IN, United States of America

Ramachandra N Rao