Jundishapur Journal of Microbiology

Published by: Kowsar

Determination of Temperature Sensitive Plasmid Copy Number in Escherichia coli by Absolute and Relative Real Time Quantitation PCR

Xianzhang Jiang 1 , Hongjiao Liu 1 , Yongchao Niu 1 , Meirong Hu 2 , Feng Qi 1 , Mingliang Zhang 1 , Jianzhong Huang 1 , * and Yong Tao 2 , *
Authors Information
1 Engineering Research Center of Industrial Microbiology, College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350117, People’s Republic of China
2 Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China
Corresponding Authors:
Article information
  • International Cardivascular Research Journal: July 2017, 10 (7); e14600
  • Published Online: June 12, 2017
  • Article Type: Research Article
  • Received: September 8, 2016
  • Revised: February 17, 2017
  • Accepted: March 9, 2017
  • DOI: 10.5812/jjm.14600

To Cite: Jiang X, Liu H, Niu Y, Hu M, Qi F, et al. Determination of Temperature Sensitive Plasmid Copy Number in Escherichia coli by Absolute and Relative Real Time Quantitation PCR, Jundishapur J Microbiol. 2017 ; 10(7):e14600. doi: 10.5812/jjm.14600.

Copyright © 2017, International Cardivascular Research Journal. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
1. Background
2. Objectives
3. Methods
4. Results
5. Discussions
  • 1. Huang H, Zheng G, Jiang W, Hu H, Lu Y. One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces. Acta Biochim Biophys Sin (Shanghai). 2015; 47(4): 231-43[DOI][PubMed]
  • 2. Chen J, Ram G, Yoong P, Penades JR, Shopsin B, Novick RP. An rpsL-based allelic exchange vector for Staphylococcus aureus. Plasmid. 2015; 79: 8-14[DOI][PubMed]
  • 3. Hoffmann F, Rinas U. Plasmid amplification in Escherichia coli after temperature upshift is impaired by induction of recombinant protein synthesis. Biotechnol Lett. 2001; 23(22): 1819-25[DOI]
  • 4. Budiharjo A, Chowdhury SP, Dietel K, Beator B, Dolgova O, Fan B, et al. Transposon mutagenesis of the plant-associated Bacillus amyloliquefaciens ssp. plantarum FZB42 revealed that the nfrA and RBAM17410 genes are involved in plant-microbe-interactions. PLoS One. 2014; 9(5)[DOI][PubMed]
  • 5. Hashimoto-Gotoh T, Yamaguchi M, Yasojima K, Tsujimura A, Wakabayashi Y, Watanabe Y. A set of temperature sensitive-replication/-segregation and temperature resistant plasmid vectors with different copy numbers and in an isogenic background (chloramphenicol, kanamycin, lacZ, repA, par, polA). Gene. 2000; 241(1): 185-91[PubMed]
  • 6. Sugimoto M, Kojima H, Tanaka A, Matsui H, Sato K, Nakamatsu T. Temperature-sensitive plasmid. 1998;
  • 7. Gruss A, Maguin E. Institut National De La Recherche Agronomique, assignee. Temperature-sensitive plasmid. 2000;
  • 8. Bramucci MG. Temperature sensitive mutant derivatives of the broad host range plasmid pBHR1. 2006;
  • 9. Lee CL, Ow DS, Oh SK. Quantitative real-time polymerase chain reaction for determination of plasmid copy number in bacteria. J Microbiol Methods. 2006; 65(2): 258-67[DOI][PubMed]
  • 10. Zhong C, Peng D, Ye W, Chai L, Qi J, Yu Z, et al. Determination of plasmid copy number reveals the total plasmid DNA amount is greater than the chromosomal DNA amount in Bacillus thuringiensis YBT-1520. PLoS One. 2011; 6(1)[DOI][PubMed]
  • 11. Weisblum B, Graham MY, Gryczan T, Dubnau D. Plasmid copy number control: isolation and characterization of high-copy-number mutants of plasmid pE194. J Bacteriol. 1979; 137(1): 635-43[PubMed]
  • 12. Projan SJ, Carleton S, Novick RP. Determination of plasmid copy number by fluorescence densitometry. Plasmid. 1983; 9(2): 182-90[PubMed]
  • 13. Genthner FJ, Hook LA, Strohl WR. Determination of the molecular mass of bacterial genomic DNA and plasmid copy number by high-pressure liquid chromatography. Appl Environ Microbiol. 1985; 50(4): 1007-13[PubMed]
  • 14. Fujimura H, Sakuma Y, Weimer T. A quantitative method for the determination of plasmid copy number in recombinant yeast. Biologicals. 1996; 24(1): 67-70[PubMed]
  • 15. Breuer S, Marzban G, Cserjan-Puschman M, Durrschmid E, Bayer K. Off-line quantitative monitoring of plasmid copy number in bacterial fermentation by capillary electrophoresis. Electrophoresis. 1998; 19(14): 2474-8[DOI][PubMed]
  • 16. Lee C, Kim J, Shin SG, Hwang S. Absolute and relative QPCR quantification of plasmid copy number in Escherichia coli. J Biotechnol. 2006; 123(3): 273-80[DOI][PubMed]
  • 17. Carapuca E, Azzoni AR, Prazeres DM, Monteiro GA, Mergulhao FJ. Time-course determination of plasmid content in eukaryotic and prokaryotic cells using real-time PCR. Mol Biotechnol. 2007; 37(2): 120-6[PubMed]
  • 18. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop R2, et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene. 1995; 166(1): 175-6[PubMed]
  • 19. Antoine R, Locht C. Isolation and molecular characterization of a novel broad-host-range plasmid from Bordetella bronchiseptica with sequence similarities to plasmids from gram-positive organisms. Mol Microbiol. 1992; 6(13): 1785-99[PubMed]
  • 20. Jiang X, Huang J, Liu H, Zhou F, Zhang M, Qi F. Fujian Normal University, assignee. The temperature sensitive plasmid pBBR1mcs-2-ts4 and uses thereof. 2015;
  • 21. Hahn FM, Eubanks LM, Testa CA, Blagg BS, Baker JA, Poulter CD. 1-Deoxy-D-xylulose 5-phosphate synthase, the gene product of open reading frame (ORF) 2816 and ORF 2895 in Rhodobacter capsulatus. J Bacteriol. 2001; 183(1): 1-11[DOI][PubMed]
  • 22. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison C3, Smith HO. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods. 2009; 6(5): 343-5[DOI][PubMed]
  • 23. Tombline G, Bartholomew LA, Urbatsch IL, Senior AE. Combined mutation of catalytic glutamate residues in the two nucleotide binding domains of P-glycoprotein generates a conformation that binds ATP and ADP tightly. J Biol Chem. 2004; 279(30): 31212-20[DOI][PubMed]
  • 24. Dhanasekaran S, Doherty TM, Kenneth J, T. B. Trials Study Group . Comparison of different standards for real-time PCR-based absolute quantification. J Immunol Methods. 2010; 354(1-2): 34-9[DOI][PubMed]
  • 25. Yu Y, Lee C, Kim J, Hwang S. Group-specific primer and probe sets to detect methanogenic communities using quantitative real-time polymerase chain reaction. Biotechnol Bioeng. 2005; 89(6): 670-9[DOI][PubMed]
  • 26. Cikos S, Bukovska A, Koppel J. Relative quantification of mRNA: comparison of methods currently used for real-time PCR data analysis. BMC Mol Biol. 2007; 8: 113[DOI][PubMed]
  • 27. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001; 25(4): 402-8[DOI][PubMed]
  • 28. Reed GF, Lynn F, Meade BD. Use of coefficient of variation in assessing variability of quantitative assays. Clin Diagn Lab Immunol. 2002; 9(6): 1235-9[PubMed]
  • 29. Kovach ME, Phillips RW, Elzer PH, Roop R2, Peterson KM. pBBR1MCS: a broad-host-range cloning vector. Biotechniques. 1994; 16(5): 800-2[PubMed]
  • 30. Elzer PH, Kovach ME, Phillips RW, Robertson GT, Peterson KM, Roop R2. In vivo and in vitro stability of the broad-host-range cloning vector pBBR1MCS in six Brucella species. Plasmid. 1995; 33(1): 51-7[DOI][PubMed]
  • 31. DeShazer D, Woods DE. Broad-host-range cloning and cassette vectors based on the R388 trimethoprim resistance gene. Biotechniques. 1996; 20(5): 762-4[PubMed]
  • 32. Obranic S, Babic F, Maravic-Vlahovicek G. Improvement of pBBR1MCS plasmids, a very useful series of broad-host-range cloning vectors. Plasmid. 2013; 70(2): 263-7[DOI][PubMed]
  • 33. Gruber S, Schwab H, Koefinger P. Versatile plasmid-based expression systems for Gram-negative bacteria--General essentials exemplified with the bacterium Ralstonia eutropha H16. N Biotechnol. 2015; 32(6): 552-8[DOI][PubMed]
  • 34. Tian M, Qu J, Bao Y, Gao J, Liu J, Wang S, et al. Construction of pTM series plasmids for gene expression in Brucella species. J Microbiol Methods. 2016; 123: 18-23[DOI][PubMed]
Creative Commons License Except where otherwise noted, this work is licensed under Creative Commons Attribution Non Commercial 4.0 International License .

Search Relations:



Create Citiation Alert
via Google Reader

Readers' Comments