Efficient transformation of the astaxanthin-producing yeast Phaffia rhodozyma.

J. Wery; J.C. Verdoes; A.J.J. van Ooyen

doi:10.1023/A:1008834600770

Efficient transformation of the astaxanthin-producing yeast Phaffia rhodozyma.

J. Wery, J.C. Verdoes, A.J.J. van Ooyen

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Abstract

An efficient transformation system for the astaxanthin-producing yeast Phaffia rhodozyma was developed based on electroporation that routinely yields approximately 1000 transformants per g of plasmid DNA. The high transformation efficiency depends on vector integration in the ribosomal DNA (rDNA) and the presence of the homologous glycolytic glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and terminator to drive the expression of the transposon Tn5 encoded kanamycin resistance gene (KmR) as a selective marker. Using this system stable transformants were obtained, carrying multiple plasmid copies. Plasmid copy number could be markedly increased by deletion of the gpd terminator from the transforming plasmid.

Original language	English
Pages (from-to)	399-405
Journal	Biotechnology techniques
Volume	12
DOIs	https://doi.org/10.1023/A:1008834600770
Publication status	Published - 1998

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title = "Efficient transformation of the astaxanthin-producing yeast Phaffia rhodozyma.",

abstract = "An efficient transformation system for the astaxanthin-producing yeast Phaffia rhodozyma was developed based on electroporation that routinely yields approximately 1000 transformants per g of plasmid DNA. The high transformation efficiency depends on vector integration in the ribosomal DNA (rDNA) and the presence of the homologous glycolytic glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and terminator to drive the expression of the transposon Tn5 encoded kanamycin resistance gene (KmR) as a selective marker. Using this system stable transformants were obtained, carrying multiple plasmid copies. Plasmid copy number could be markedly increased by deletion of the gpd terminator from the transforming plasmid.",

author = "J. Wery and J.C. Verdoes and {van Ooyen}, A.J.J.",

year = "1998",

doi = "10.1023/A:1008834600770",

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T1 - Efficient transformation of the astaxanthin-producing yeast Phaffia rhodozyma.

AU - Wery, J.

AU - Verdoes, J.C.

AU - van Ooyen, A.J.J.

PY - 1998

Y1 - 1998

N2 - An efficient transformation system for the astaxanthin-producing yeast Phaffia rhodozyma was developed based on electroporation that routinely yields approximately 1000 transformants per g of plasmid DNA. The high transformation efficiency depends on vector integration in the ribosomal DNA (rDNA) and the presence of the homologous glycolytic glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and terminator to drive the expression of the transposon Tn5 encoded kanamycin resistance gene (KmR) as a selective marker. Using this system stable transformants were obtained, carrying multiple plasmid copies. Plasmid copy number could be markedly increased by deletion of the gpd terminator from the transforming plasmid.

AB - An efficient transformation system for the astaxanthin-producing yeast Phaffia rhodozyma was developed based on electroporation that routinely yields approximately 1000 transformants per g of plasmid DNA. The high transformation efficiency depends on vector integration in the ribosomal DNA (rDNA) and the presence of the homologous glycolytic glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and terminator to drive the expression of the transposon Tn5 encoded kanamycin resistance gene (KmR) as a selective marker. Using this system stable transformants were obtained, carrying multiple plasmid copies. Plasmid copy number could be markedly increased by deletion of the gpd terminator from the transforming plasmid.

U2 - 10.1023/A:1008834600770

DO - 10.1023/A:1008834600770

M3 - Article

VL - 12

SP - 399

EP - 405

JO - Biotechnology techniques

JF - Biotechnology techniques

SN - 0951-208X

ER -

Efficient transformation of the astaxanthin-producing yeast Phaffia rhodozyma.

Abstract

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