Autonomously replicating linear plasmids that facilitate the analysis of replication origin function in Candida albicans

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Autonomously replicating linear plasmids that facilitate the analysis of replication origin function in Candida albicans. / Bijlani, Swati; Thevandavakkam, Mathuravani A.; Berman, Judith; Tsai, Hung-Ji.

In: mSphere, Vol. 4, No. 2, 06.03.2019, p. e00103-19.

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Bijlani, Swati ; Thevandavakkam, Mathuravani A. ; Berman, Judith ; Tsai, Hung-Ji. / Autonomously replicating linear plasmids that facilitate the analysis of replication origin function in Candida albicans. In: mSphere. 2019 ; Vol. 4, No. 2. pp. e00103-19.

Bibtex

@article{abfe1d996f4e4c3cb60d7da60afa2313,
title = "Autonomously replicating linear plasmids that facilitate the analysis of replication origin function in Candida albicans",
abstract = "The ability to generate autonomously replicating plasmids has been elusive in Candida albicans, a prevalent human fungal commensal and pathogen. Instead, plasmids generally integrate into the genome. Here, we assessed plasmid and transformant properties, including plasmid geometry, transformant colony size, four selectable markers, and potential origins of replication, for their ability to drive autonomous plasmid maintenance. Importantly, linear plasmids with terminal telomere repeats yielded many more autonomous transformants than circular plasmids with the identical sequences. Furthermore, we could distinguish (by colony size) transient, autonomously replicating, and chromosomally integrated transformants (tiny, medium, and large, respectively). Candida albicans URA3 and a heterologous marker, ARG4, yielded many transient transformants indicative of weak origin activity; the replication of the plasmid carrying the heterologous LEU2 marker was highly dependent upon the addition of a bona fide origin sequence. Several bona fide chromosomal origins, with an origin fragment of ~100 bp as well as a heterologous origin, panARS, from Kluyveromyces lactis, drove autonomous replication, yielding moderate transformation efficiency and plasmid stability. Thus, C. albicans maintains linear plasmids that yield high transformation efficiency and are maintained autonomously in an origin-dependent manner.",
keywords = "CaURA3, Linear plasmids, Replication, Replication origins, Telomere repeats",
author = "Swati Bijlani and Thevandavakkam, {Mathuravani A.} and Judith Berman and Hung-Ji Tsai",
year = "2019",
month = mar,
day = "6",
doi = "10.1128/mSphere.00103-19",
language = "English",
volume = "4",
pages = "e00103--19",
journal = "mSphere",
issn = "2379-5042",
publisher = "American Society for Microbiology",
number = "2",

}

RIS

TY - JOUR

T1 - Autonomously replicating linear plasmids that facilitate the analysis of replication origin function in Candida albicans

AU - Bijlani, Swati

AU - Thevandavakkam, Mathuravani A.

AU - Berman, Judith

AU - Tsai, Hung-Ji

PY - 2019/3/6

Y1 - 2019/3/6

N2 - The ability to generate autonomously replicating plasmids has been elusive in Candida albicans, a prevalent human fungal commensal and pathogen. Instead, plasmids generally integrate into the genome. Here, we assessed plasmid and transformant properties, including plasmid geometry, transformant colony size, four selectable markers, and potential origins of replication, for their ability to drive autonomous plasmid maintenance. Importantly, linear plasmids with terminal telomere repeats yielded many more autonomous transformants than circular plasmids with the identical sequences. Furthermore, we could distinguish (by colony size) transient, autonomously replicating, and chromosomally integrated transformants (tiny, medium, and large, respectively). Candida albicans URA3 and a heterologous marker, ARG4, yielded many transient transformants indicative of weak origin activity; the replication of the plasmid carrying the heterologous LEU2 marker was highly dependent upon the addition of a bona fide origin sequence. Several bona fide chromosomal origins, with an origin fragment of ~100 bp as well as a heterologous origin, panARS, from Kluyveromyces lactis, drove autonomous replication, yielding moderate transformation efficiency and plasmid stability. Thus, C. albicans maintains linear plasmids that yield high transformation efficiency and are maintained autonomously in an origin-dependent manner.

AB - The ability to generate autonomously replicating plasmids has been elusive in Candida albicans, a prevalent human fungal commensal and pathogen. Instead, plasmids generally integrate into the genome. Here, we assessed plasmid and transformant properties, including plasmid geometry, transformant colony size, four selectable markers, and potential origins of replication, for their ability to drive autonomous plasmid maintenance. Importantly, linear plasmids with terminal telomere repeats yielded many more autonomous transformants than circular plasmids with the identical sequences. Furthermore, we could distinguish (by colony size) transient, autonomously replicating, and chromosomally integrated transformants (tiny, medium, and large, respectively). Candida albicans URA3 and a heterologous marker, ARG4, yielded many transient transformants indicative of weak origin activity; the replication of the plasmid carrying the heterologous LEU2 marker was highly dependent upon the addition of a bona fide origin sequence. Several bona fide chromosomal origins, with an origin fragment of ~100 bp as well as a heterologous origin, panARS, from Kluyveromyces lactis, drove autonomous replication, yielding moderate transformation efficiency and plasmid stability. Thus, C. albicans maintains linear plasmids that yield high transformation efficiency and are maintained autonomously in an origin-dependent manner.

KW - CaURA3

KW - Linear plasmids

KW - Replication

KW - Replication origins

KW - Telomere repeats

UR - http://www.scopus.com/inward/record.url?scp=85062628104&partnerID=8YFLogxK

U2 - 10.1128/mSphere.00103-19

DO - 10.1128/mSphere.00103-19

M3 - Article

C2 - 30842269

AN - SCOPUS:85062628104

VL - 4

SP - e00103-19

JO - mSphere

JF - mSphere

SN - 2379-5042

IS - 2

ER -