Characterization of plasmid burden and copy number in Saccharomyces cerevisiae for optimization of metabolic engineering applications.
Characterization of plasmid burden and copy number in Saccharomyces cerevisiae for optimization of metabolic engineering applications.
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DOI：
10.1111/1567-1364.12016
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发表时间：
2013-02
影响因子：
3.2
通讯作者：
Alper HS
中科院分区：
文献类型：
--
作者：
Karim AS;Curran KA;Alper HS
许多代谢工程和酵母中的基因工程应用都取决于质粒的使用，尽管使用了质粒，并且有可能使用的潜水员收集，但基本上缺乏对常用的DNA质粒的影响质粒成分可以影响质粒载荷和伯嫩，我们表征了通常在酵母中使用的2μ和centromeric质粒的主要属性选择标记物，启动子，复制的起源和应变倍质的影响对授予的生长速率和质粒拷贝数的影响。选择标记，尤其是当利用质粒的质粒燃烧时，传统上归因于质粒的复制和维持质粒DNA在单倍体酵母中的作用较小，但在二倍体菌株也可以显着改变质粒拷贝数。工程应用。
Many metabolic engineering and genetic engineering applications in yeast rely on the use of plasmids. Despite their pervasive use and the diverse collections available, there is a fundamental lack of understanding of how commonly used DNA plasmids affect the cell’s ability to grow and how the choice of plasmid components can influence plasmid load and burden. In this study, we characterized the major attributes of the 2μ and centromeric plasmids typically used in yeast by examining the impact of choice of selection marker, promoter, origin of replication, and strain ploidy on conferred growth rates and plasmid copy number. We conclude that the “plasmid burden,” as demonstrated by a reduced growth rate, is primarily due to the choice of selection marker, especially when auxotrophic markers are utilized. The plasmid burden traditionally attributed to replication and maintenance of plasmid DNA plays only a minor role in haploid yeast yet is much more significant in diploid strains. The selection marker can also significantly change plasmid copy number. In fact, plasmid copy number can be influenced to some extent by all of the parameters tested. The information presented in this study will allow for more rational design and selection of plasmids for engineering applications.
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DOI：
10.1073/pnas.75.4.1929
发表时间：
1978-01-01
影响因子：
11.1
作者：
HINNEN, A;HICKS, JB;FINK, GR
通讯作者：
FINK, GR
影响因子：
3.2
作者：
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通讯作者：
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影响因子：
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作者：
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影响因子：
3.3
作者：
Hu, X. H.;Wang, M. H.;Luo, Z. W.
通讯作者：
Luo, Z. W.
影响因子：
64.8
作者：
Dymond, Jessica S.;Richardson, Sarah M.;Coombes, Candice E.;Babatz, Timothy;Muller, Heloise;Annaluru, Narayana;Blake, William J.;Schwerzmann, Joy W.;Dai, Junbiao;Lindstrom, Derek L.;Boeke, Annabel C.;Gottschling, Daniel E.;Chandrasegaran, Srinivasan;Bader, Joel S.;Boeke, Jef D.
通讯作者：
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