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US 6,284,534 B1Granted
Yeast vector comprising a shortened promoter sequence
AssigneeKirin Beer Kabushiki Kaisha
InventorsKeiji Kondo, Yutaka Miura
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AssigneeKirin Beer Kabushiki Kaisha
InventorsKeiji Kondo, Yutaka Miura
Estimating the term…
An object of the present invention is to provide a vector which can be integrated into a yeast chromosome in a high number of copies. Another object of the present invention is to provide a modified vector which can be integrated into the yeast chromosome in a high number of copies and of which expression units stably maintain on the chromosome. The vector according to the present invention comprises a marker gene for selecting transformants, a shortened promoter sequence which is operably linked to the marker gene and a sequence homologous to the chromosomal DNA of Candida utilis, and optionally a heterologous gene or a gene derived from C. utilis, wherein the vector is linearized by cleaving within said homologous DNA sequence or at both ends of the homologous DNA sequence with restriction enzymes, and wherein the heterologous gene or the gene derived from C. utilis can be integrated into the chromosomal DNA of C. utilis by homologous recombination.
A vector comprising a marker gene for selecting transformants, a shortened promoter sequence that is operably linked to the marker gene, and a homologous DNA sequence that is homologous to chromosomal DNA, other than an rDNA sequence, of Candid utilis, and optionally a heterologous gene or gene isolated from C. utilis, wherein the vector is linearized by cleaving within or at both ends of the homologous DNA sequence with a restriction enzyme, wherein the heterologous gene or gene isolated from Candida utilis can be integrated into the chromosomal DNA of Candida utilis by homologous recombination, and wherein the shortened promoter is a promoter truncated at its 5′ terminus to decrease, relative to pCLRE11, the transformation frequency of the vector in host cells of Candida utilis and to increase, relative to pCLRE11, copy number of the vector in host cells of Candida utilis.
Dependencies are inferred from the claim text.