Promoters from chlorella virus genes providing for expression of genes in prokaryotic and eukaryotic hosts
AssigneeBoard of Regents, University of Nebraska-Lincoln
InventorsAmitava Mitra, James L. Van Etten
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Abstract
The invention is directed to novel promoters or mutants thereof from Chlorella virus DNA methyltansferase genes. A Chlorella virus gene promoter is operably linked to a first and/or second DNA sequence encoding a gene that is different from the Chlorella virus to form an expression cassette. An expression cassette can be introduced into prokaryotic and/or eukaryotic cells and can provide for a high level of expression of the gene encoded by the first and/or second DNA sequence. The invention also provides a method for screening other Chlorella virus genes for promoters that can function to express a heterologous gene in prokaryotic and/or eukaryotic hosts.
Claims22
A method of identifying whether a DNA sequence or mutants thereof substantially corresponding to a Chlorella virus genome sequence can function to promote expression of one or more heterologous gene, comprising the steps of:
- (a) constructing a plasmid vector comprising a promoter or a mutant thereof substantially corresponding to a promoter from a Chlorella virus DNA methyltransferase gene operably linked to a first DNA sequence encoding the one or more heterologous gene that is different from the Chlorella virus DNA methyltransferase gene;
- (b) transforming said plasmid vector into host cells selected from the group consisting of prokaryotic host cells and eukaryotic host cells;
- and
- (c) detecting expression of one or more of said heterologous gene or genes.
A method of identifying whether a DNA sequence or mutants thereof substantially corresponding to a Chlorella virus genome sequence can function to promote expression of one or more heterologous gene, comprising the steps of:
- (a) constructing a plasmid vector comprising a promoter or a mutant thereof substantially corresponding to a promoter from a Chlorella virus DNA methyltransferase gene operably linked to a first DNA sequence encoding the one or more heterologous gene that is different from the Chlorella virus DNA methyltransferase gene and a 3′ nontranslated regulatory DNA sequence operably linked to the first DNA sequence;
- (c) transforming said plasmid vector into host cells selected from the group consisting of prokaryotic host cells and eukaryotic host cells;
- and
- (c) detecting expression of one or more of said heterologous gene or genes.
A method of identifying whether a DNA sequence or mutants thereof substantially corresponding to a Chlorella virus genome sequence can function to promote expression of one or more heterologous gene, comprising the steps of:
- (a) constructing a plasmid vector comprising a promoter or a mutant thereof substantially corresponding to a promoter from a Chlorella virus DNA methyltransferase gene operably linked to a first DNA sequence encoding the one or more heterologous gene that is different from the Chlorella virus DNA methyltransferase gene and a 3′ nontranslated regulatory DNA sequence operably linked to the first DNA sequence, wherein the 3′ nontranslated regulatory DNA sequence has a polyadenylation sequence suitable for functioning in plant cells;
- (d) transforming said plasmid vector into host cells selected from the group consisting of prokaryotic host cells and eukaryotic host cells;
- and
- (c) detecting expression of one or more of said heterologous gene or genes.
Dependencies are inferred from the claim text.