COSPLAY: An expandable toolbox for combinatorial and swift generation of expression plasmids in yeast

被引:5
作者
Goulev, Youlian [1 ,2 ,3 ,4 ,5 ]
Matifas, Audrey [1 ,2 ,3 ,4 ]
Heyer, Vincent [1 ,2 ,3 ,4 ]
Reina-San-Martin, Bernardo [1 ,2 ,3 ,4 ]
Charvin, Gilles [1 ,2 ,3 ,4 ]
机构
[1] Inst Genet & Biol Mol & Cellulaire, Dept Dev Biol & Stem Cells, Illkirch Graffenstaden, France
[2] CNRS, UMR7104, Illkirch Graffenstaden, France
[3] INSERM, U964, Illkirch Graffenstaden, France
[4] Univ Strasbourg, Illkirch Graffenstaden, France
[5] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA
关键词
NEAREST-NEIGHBOR THERMODYNAMICS; CENTER-DOT-C; SACCHAROMYCES-CEREVISIAE; SHUTTLE VECTORS; DNA; MISMATCHES; CLONING; DELETION; NMR; PCR;
D O I
10.1371/journal.pone.0220694
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A large number of genetic studies in yeast rely on the use of expression vectors. To facilitate the experimental approach of these studies, several collections of expression vectors have been generated (YXplac, pRS series, etc.). Subsequently, these collections have been expanded by adding more diversity to many of the plasmid features, including new selection markers and new promoter sequences. However, the ever growing number of plasmid features makes it unrealistic for research labs to maintain an up-to-date collection of plasmids. Here, we developed the COSPLAY toolbox: a Golden Gate approach based on the scheme of a simple modular plasmid that recapitulates and completes all the properties of the pRS plasmids. The COSPLAY toolbox contains a basal collection of individual functional modules. Moreover, we standardized a simple and rapid, software-assisted protocol which facilitates the addition of new personalized modules. Finally, our toolbox includes the possibility to select a genomic target location and to perform a single copy integration of the expression vector.
引用
收藏
页数:18
相关论文
共 44 条
[1]   A suite of Gateway® cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae [J].
Alberti, Simon ;
Gitler, Aaron D. ;
Lindquist, Susan .
YEAST, 2007, 24 (10) :913-919
[2]   Nearest-neighbor thermodynamics of internal A•C mismatches in DNA:: Sequence dependence and pH effects [J].
Allawi, HT ;
SantaLucia, J .
BIOCHEMISTRY, 1998, 37 (26) :9435-9444
[3]   Nearest neighbor thermodynamic parameters for internal G•A mismatches in DNA [J].
Allawi, HT ;
SantaLucia, J .
BIOCHEMISTRY, 1998, 37 (08) :2170-2179
[4]   Thermodynamics and NMR of internal GT mismatches in DNA [J].
Allawi, HT ;
SantaLucia, J .
BIOCHEMISTRY, 1997, 36 (34) :10581-10594
[5]   Thermodynamics of internal C•T mismatches in DNA [J].
Allawi, HT ;
Santalucia, J .
NUCLEIC ACIDS RESEARCH, 1998, 26 (11) :2694-2701
[6]   NMR solution structure of a DNA dodecamer containing single G•T mismatches [J].
Allawi, HT ;
SantaLucia, J .
NUCLEIC ACIDS RESEARCH, 1998, 26 (21) :4925-4934
[7]   Controlling promoter strength and regulation in Saccharomyces cerevisiae using synthetic hybrid promoters [J].
Blazeck, John ;
Garg, Rishi ;
Reed, Ben ;
Alper, Hal S. .
BIOTECHNOLOGY AND BIOENGINEERING, 2012, 109 (11) :2884-2895
[8]  
Brachmann CB, 1998, YEAST, V14, P115
[9]   Protein sequestration generates a flexible ultrasensitive response in a genetic network [J].
Buchler, Nicolas E. ;
Cross, Frederick R. .
MOLECULAR SYSTEMS BIOLOGY, 2009, 5
[10]   New and Redesigned pRS Plasmid Shuttle Vectors for Genetic Manipulation of Saccharomyces cerevisiae [J].
Chee, Mark K. ;
Haase, Steven B. .
G3-GENES GENOMES GENETICS, 2012, 2 (05) :515-526