Part:BBa_K658020

iccdB->GFP

This device is designed to test how a population-control device could influence the expression of its downstream genes.

This part is made up of two subparts, a population-control device iccdB0.6 (BBa_K658003) and an EGFP (BBa_J04630) reporter. The population-control device iccdB0.6 (BBa_K658003) has the ability to maintain steady-state cell density at a relatively low value and extend the steady state in cell growth. The reporter EGFP’s (BBa_J04630) expression is regulated by the iccdB0.6 in upstream.

Figure 1 Proposed mechanism of iccdB0.6-GFP.

Description

This device is an example of how population-control device could be used in regulating the expression of our desired product, such as antibiotics and proteins.

Result

Figure 2 Experimentally measured fluorescence at steady state of BL21's cells with iccdB0.6-GFP(BBa_K658020) and BBa_K658021.

Reference

[1]You L, Cox RS, Weiss R, Arnold FH. Programmed population control by cell-cell communication and regulated killing[J]. Nature, 2004, 428(6985): 868-871.

[2]Koch B, Liljefors T, Persson T , Nielsen J, Kjelleberg S, Givskov M. The LuxR receptor: the sites of interaction with quorum-sensing signals and inhibitors[J].Microbiology , 2005, 151 (11): 3589-3602.

[3] Kumari A, Pasini P, Deo S. K., Flomenhoft D, Shashidhar S, Daunert S. Biosensing Systems for the Detection of Bacterial Quorum Signaling Molecules[J]. Analytical Chemistry,2005, 78 (22): 7603–7609.

[4] Eberhard A, Burlingame A. L, Eberhard C, Kenyon G. L, Nealson K. H,Oppenheimer N. J. Structural identification of autoinducer of Photobacterium fischeri luciferase[J]. Biochemistry ,1981,20 (9): 2444–2449.

[5] http://scholarworks.umass.edu/dissertations/AAI3397713/

[6] Gerdes K. Toxin-antitoxin modules may regulate synthesis of macromolecules during nutritional stress[J]. J. Bacteriol,2000,182 (3): 561–721.

[7]Philippe B,Martine C.Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes[J].Molecular Biology,1992,226(3):735-745.

[8]Devine J.H,Shadel G.S,Baldwin T.O. Identification of the operator of the lux regulon from the Vibrio fischeri strain ATCC7744. Proceedings of the National Academy of Sciences.1989.86(15):5688-5692.

[9] http://www.uniprot.org/uniprot/P12747

[10]Miki T,Yoshioka K,Horiuchi T. Control of cell division by sex factor F in Escherichia coli. I. The 42.84-43.6 F segment couples cell division of the host bacteria with replication of plasmid DNA. Molecular Biology,1984,177(4):605-625.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 718
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2347
  Illegal BsaI.rc site found at 1874
  Illegal BsaI.rc site found at 3310