Cetus Corporation

James H Meade

David H Gelfand

Hing C Wong

Michael A Innis

Arie Ben-Bassat

Shing Chang

Rony Tal

Anna Lisa Fear

Moshe Benziman

Roger D Calhoon

Nucleic acid sequences encoding the bacterial cellulose synthase operon derived from Acetobacter are disclosed. Methods for isolating the genes, vectors containing the genes, and transformed hosts useful for the expression of recombinant bacterial cellulose synthase or production of cellulose are also described.

Methods and nucleic acid sequences for the expression of the cellulose

A gene having a DNA sequence complementary to that of the glucoamylase polypeptide mRNA from a fungal species, preferably Aspergillus awamori, is prepared. The mRNA is an approximately 2.2 kilobase poly A RNA obtained from fungal cells grown under conditions of glucoamylase induction. Reverse transcription of the mRNA provides a glucoamylase probe used to identify genomic digest fragments containing glucoamylase gene regions, which are sequenced to locate the introns and exons. The genomic fragments are spliced together to form a gene having a DNA sequence with altered or deleted introns which codes for fungal glucoamylase protein and is capable, when correctly combined with a cleaved DNA expression vector, of expressing a non-native protein having glucoamylase enzyme activity upon transformation of a host organism by the vector. The host is preferably bacteria or yeast. The transformed yeast host may be used to produce ethanol.

DNA expression vector and use thereof

Chimeric plasmids capable of transforming bacteria and yeast are described. The plasmids carry the Cm (chloramphenicol resistance) gene and the Tc (tetracycline resistance) gene as selectable markers. The Cm gene allows the plasmids to be selected for in wild-type strains of the yeast Saccharomyces cerevisiae. The Tc gene allows heterologous genes cloned into the plasmids to be selected for in Escherichia coli bacteria.

Chimeric plasmids that replicate in bacteria and yeast and

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Pinole, CA, United States of America

James H Meade