Rapid determination of plasmid copy number

Schmidt T, Friehs K, Flaschel E (1996)
JOURNAL OF BIOTECHNOLOGY 49(1-3): 219-229.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
Plasmid copy number, the number of expression vectors per host cell, is a key variable in recombinant microbial cultivation. Therefore, it would be very helpful, if the plasmid copy number could be determined during the operating process period. A rapid quantification of this important process variable would even open the possibility of its use in process control. However, current assays like gel electrophoresis, CsCl-gradient centrifugation, HPLC and other methods are time consuming and difficult to quantify. Indirect methods: like the correlation of copy number with, e.g. the activity of an enzyme, coded on the plasmid, are prone to errors due to the production kinetics, turnover rate and protein denaturation. Here, a method is presented, which enables the plasmid copy number to be determined in less than 30 min. This novel procedure based on plasmid isolation by means of a commercial DNA-isolation kit and quantification by capillary electrophoresis, should allow the copy number to be used in process control.
Erscheinungsjahr
Zeitschriftentitel
JOURNAL OF BIOTECHNOLOGY
Band
49
Ausgabe
1-3
Seite(n)
219-229
ISSN
PUB-ID

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Schmidt T, Friehs K, Flaschel E. Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY. 1996;49(1-3):219-229.
Schmidt, T., Friehs, K., & Flaschel, E. (1996). Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY, 49(1-3), 219-229. doi:10.1016/0168-1656(96)01519-2
Schmidt, T., Friehs, K., and Flaschel, E. (1996). Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY 49, 219-229.
Schmidt, T., Friehs, K., & Flaschel, E., 1996. Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY, 49(1-3), p 219-229.
T. Schmidt, K. Friehs, and E. Flaschel, “Rapid determination of plasmid copy number”, JOURNAL OF BIOTECHNOLOGY, vol. 49, 1996, pp. 219-229.
Schmidt, T., Friehs, K., Flaschel, E.: Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY. 49, 219-229 (1996).
Schmidt, T, Friehs, Karl, and Flaschel, Erwin. “Rapid determination of plasmid copy number”. JOURNAL OF BIOTECHNOLOGY 49.1-3 (1996): 219-229.

14 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Quantification of Plasmid Copy Number with Single Colour Droplet Digital PCR.
Plotka M, Wozniak M, Kaczorowski T., PLoS One 12(1), 2017
PMID: 28085908
Capacitive biosensor for quantification of trace amounts of DNA.
Numnuam A, Kanatharana P, Mattiasson B, Asawatreratanakul P, Wongkittisuksa B, Limsakul C, Thavarungkul P., Biosens Bioelectron 24(8), 2009
PMID: 19201186
Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes.
Skulj M, Okrslar V, Jalen S, Jevsevar S, Slanc P, Strukelj B, Menart V., Microb Cell Fact 7(), 2008
PMID: 18328094
The copy-number of plasmids and other genetic elements can be determined by SYBR-Green-based quantitative real-time PCR.
Providenti MA, O'Brien JM, Ewing RJ, Paterson ES, Smith ML., J Microbiol Methods 65(3), 2006
PMID: 16216354
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Jana S, Deb JK., Appl Microbiol Biotechnol 67(3), 2005
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Cheng L, Li TY, Zhang Y., J Biochem Mol Biol 37(3), 2004
PMID: 15469718
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Noites IS, O'Kennedy RD, Levy MS, Abidi N, Keshavarz-Moore E., Biotechnol Bioeng 66(3), 1999
PMID: 10577474
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PMID: 10527521
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Breuer S, Marzban G, Cserjan-Puschman M, Dürrschmid E, Bayer K., Electrophoresis 19(14), 1998
PMID: 9820970

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