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Cloning and sequencing of the URA3 gene of Kluyveromyces marxianus CBS 6556
Author(s) -
Bergkamp Ronald J. M.,
Geerse Ruud H.,
Planta Rudi J.,
Verbakel John M. A.
Publication year - 1993
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320090614
Subject(s) - ura3 , kluyveromyces marxianus , biology , kluyveromyces lactis , gene , kluyveromyces , mutant , genomic dna , microbiology and biotechnology , genetics , genomic library , saccharomyces cerevisiae , biochemistry , peptide sequence
The URA3 gene, coding for orotidine‐5′‐phosphate decarboxylase, from Kluyveromyces marxianus CBS 6556, was isolated from a genomic DNA library. The K. marxianus URA3 gene encodes a protein of 267 amino acids with a calculated molecular weight of 29·3 kDa. Comparison of the K. marxianus protein with the corresponding enzymes of Saccharomyces cerevisiae and Kluyveromyces lactis showed amino acid sequence identifies of 81% and 88%, respectively. Using contour‐clamped homogeneous electric field gel electrophoresis, the genomic copy was found to be located on chromosome VI. We have used the cloned gene for the construction of a K. marxianus leu2 mutant. This mutant contains no heterologous sequences, which is essential to make it acceptable for application in the food industry.