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AssigneeWisconsin Alumni Research Foundation
InventorsWaclaw T. Szybalski, Jadwiga Wild, Zdenka Hradecna
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The present invention provides an expression vector adapted for inducible amplification and inducible transcription of a heterologous polynucleotide operably linked to a transcriptional promoter provided in the vector. The invention further provides a method for using the vector to produce large quantities of a heterologous polypeptide encoded by the heterologous polynucleotide. The pBAC of the present invention is derived from a BAC cloning vector by modifying the BAC cloning vector to include a conditional ori for amplifying the pBAC and an inducible promoter for regulating the transcription of an inserted gene. Both the conditional ori and the inducible promoter can be activated jointly or separately by suitable signals in a host cell. Once a polynucleotide of interest is inserted into the pBAC and the pBAC is introduced into a host cell, the level of the product encoded by the polynucleotide can be controlled by regulating the copy number of the expression vector or the activity of the promoter under which transcription of the inserted polynucleotide is controlled, or both.
An expression vector for transcribing in a host cell a heterologous polynucleotide that encodes a heterologous polypeptide, the expression vector comprising:
A bacterial artificial chromosome for transcribing in a host cell a heterologous polynucleotide that encodes a heterologous polypeptide, the bacterial artificial chromosome, comprising:
A host cell, comprising in its interior:
A method for inducing expression of a heterologous polypeptide in a host cell that comprises an expression vector, the expression vector comprising an origin of replication responsive to a replication-inducing agent and a transcriptional promoter responsive to a transcription-inducing agent and operably linked upstream of a heterologous polynucleotide that encodes the heterologous polypeptide, the heterologous polypeptide being toxic to the host cell, the method comprising the steps of:
An expression vector for transcribing a heterologous polynucleotide that encodes a heterologous polypeptide, the expression vector comprising:
A bacterial artificial chromosome for transcribing a heterologous polynucleotide that encodes a heterologous polypeptide, the bacterial artificial chromosome, comprising:
An expression vector for transcribing a heterologous polynucleotide that encodes a heterologous polypeptide, the expression vector comprising:
A bacterial artificial chromosome for transcribing a heterologous polynucleotide that encodes a heterologous polypeptide, the bacterial artificial chromosome, comprising:
Dependencies are inferred from the claim text.