Plasmids for in vivo construction of integrative Candida albicans vectors in Saccharomyces cerevisiae

Neide Vieira; Filipa Pereira; Margarida Casal; Alistair J.P. Brown; Sandra Paiva; Björn Johansson

doi:10.1002/yea.1800

Plasmids for in vivo construction of integrative Candida albicans vectors in Saccharomyces cerevisiae

Neide Vieira, Filipa Pereira, Margarida Casal, Alistair J.P. Brown, Sandra Paiva, Björn Johansson^*

^*Corresponding author for this work

Medical Sciences

University of Minho

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

A general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli, possibly because of their toxic effects. The ligation of S. cerevisiae 2 μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2 μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors, CIp10, CIp20 and CIp30, are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nos CIp10-2 μ (GU550119), CIp20-2 μ (GU550120) and CIp30-2 μ (GU550121).

Original language	English
Pages (from-to)	933-939
Number of pages	7
Journal	Yeast
Volume	27
Issue number	11
DOIs	https://doi.org/10.1002/yea.1800
Publication status	Published - 1 Nov 2010

Keywords

Candida albicans
Gap-repair
in vivo
Plasmid integration
Toxic genes
Yeast

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title = "Plasmids for in vivo construction of integrative Candida albicans vectors in Saccharomyces cerevisiae",

abstract = "A general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli, possibly because of their toxic effects. The ligation of S. cerevisiae 2 μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2 μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors, CIp10, CIp20 and CIp30, are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nos CIp10-2 μ (GU550119), CIp20-2 μ (GU550120) and CIp30-2 μ (GU550121).",

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AU - Paiva, Sandra

AU - Johansson, Björn

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N2 - A general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli, possibly because of their toxic effects. The ligation of S. cerevisiae 2 μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2 μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors, CIp10, CIp20 and CIp30, are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nos CIp10-2 μ (GU550119), CIp20-2 μ (GU550120) and CIp30-2 μ (GU550121).

AB - A general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli, possibly because of their toxic effects. The ligation of S. cerevisiae 2 μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2 μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors, CIp10, CIp20 and CIp30, are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nos CIp10-2 μ (GU550119), CIp20-2 μ (GU550120) and CIp30-2 μ (GU550121).

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Plasmids for in vivo construction of integrative Candida albicans vectors in Saccharomyces cerevisiae

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