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Chapter 29 : Plasmids as Tools for Containment

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Abstract:

This chapter reviews and discusses different aspects of containment, highlighting those systems devoted to contain organisms that remove toxic pollutants. There are two main strategies to diminish the potential risks associated with the deliberate or unintentional release of genetically modified organisms (GMOs) into the open environment. The lethal functions most extensively used for developing active containment circuits are those that disrupt the membrane potential, specially the two-component toxin-antidote systems involved in post-segregational killing of plasmid-free cells and their chromosomal counterparts. Biological containment systems have been engineered on plasmids using the Plac promoter and the Lacl repressor from , and they are triggered by addition of isopropyl-β-D-thiogalactopyranoside (IPTG). The most advanced containment systems are those developed for bacteria that degrade pollutants. Although the biological containment systems increase the predictability of GMOs, one of the main concerns about the release of such GMOs to the environment is how recombinant DNA can spread among indigenous bacterial populations. Lethal donation circuits, such as those described for gene containment, constitute interesting tools to explore the ecological and evolutionary consequences of shifting the natural equilibrium between genetic change and genetic constancy toward the latter. Many lethal functions and regulatory circuits have been used and combined to design efficient containment systems. Active containment systems are a major tool to reduce the uncertainty associated with the introduction of monocultures, genetically engineered or not, into target habitats.

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
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Figures

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Figure 1

Concept of biological and gene containment. (A) In biological containment the GMO is restricted to the target habitat for a limited time. To accomplish this, the organism is engineered with a suicide circuit (●) that usually is switched off (survival), but it becomes activated ( ) in response to a specific environmental signal, leading to cell death. (B) In gene containment, it is the recombinant DNA rather than the organism itself that is the subject of containment. To create a barrier that restricts dispersal of such novel DNA from the GMO to the indigenous microbiota, a genetic circuit based on a lethal function closely linked to the recombinant DNA (●) needs to be engineered in a way that the lethal function becomes activated ( ) in the potential recipients of the contained DNA, leading to the death of such cells.

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
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Image of Figure 2
Figure 2

Schematic representations of molecular mechanisms for active containment systems. (A) A control element responds to the appropriate environmental signal through a sensor protein ( ) and a cognate regulated promoter (Preg) and regulates at the transcriptional level the expression of a lethal function (■). (B) The control element can be engineered as a double transcriptional regulatory circuit. The sensor protein ( ) recognizes an environmental signal and interacts with the cognate regulated promoter (Preg1), which, in turn, drives transcription of a second regulator ( ) that controls the expression of the Preg2 promoter running transcription of the lethal gene (■). (C) The control element may involve a posttranslational regulation through an immunity protein ( ) that specifically neutralizes the killing effect of the constitutively expressed lethal function ( ). To this end, the expression of the immunity gene is driven by the Preg promoter under control of the sensor protein ( ) that responds to the environmental signal.

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
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Figure 3

Rationale of a model biological containment system for biodegraders. The control element consists of a double regulatory circuit based on (i) the XylS sensor protein that recognizes benzoate or benzoate analogues (alkyl- and halo-benzoates) and stimulates gene expression from the Pm promoter; and (ii) the lael gene, whose expression is driven by the Pm promoter, coding for the Lael repressor protein that inhibits the PA1/04/03 promoter (P*). The expression of the lethal gene is under control of the PA1/04/03(P*) promoter. (A) In the presence of benzoate or benzoate analogues the control element is switched on and the lethal function is not produced (survival). XylSa, active conformation of XylS protein. (B) Once bacteria complete the degradation of the aromatic compound or they spread to a noil polluted site, the control system is switched off and, as a consequence, the lethal function is produced (cell death). XylS1, inactive conformation of XylS protein.

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
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Figure 4

Rationale of a model plasmid containment system. The scheme has been modified from reference 10. Gene containment is achieved by a lethal donation of a killing function. In the GMO (donor cell), the lethal gene is closely linked to the novel trait in a plasmid, and the toxic effect of the lethal function is neutralized (Lethal1) by the product of an immunity gene (1mm) located at the chromosome, such that cotransfer of the lethal and immunity functions will be an extremely low-frequency event. Plasmid transfer to a nonimmune organism will lead to the activation of the lethal function (LethalA) and to the rapid killing of the recipient cells, thus preventing the spread of the plasmid and the associated novel trait.

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
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References

/content/book/10.1128/9781555817732.chap29
1. Ahrenholtz, I.,, M. G. Lorenz,, and W. Wackernagel. 1994. A conditional suicide system in Escherichia coli based on the intracellular degradation of DNA. Appl. Environ. Microbiol. 60: 3746 3751.
2. Bej, A. K.,, M. H. Perlin,, and R. M. Atlas. 1988. Model suicide vector for containment of genetically engineered microorganisms. Appl. Environ. Microbiol. 54: 2472 2477.
3. Boynton, Z. L.,, J. J. Koon,, E. M. Brennan,, J. D. Clouart,, D. M. Horowitz,, T. U. Gerngross,, and G. W. Huisman. 1999. Reduction of cell lysate viscosity during processing of poly(3- hydroxyalkanoates) by chromosomal integration of the staphylococcal nuclease gene in Pseudomonas putida. Appl. Environ. Microbiol. 65: 1524 1529.
4. Clerc, S.,, and P. Simonet. 1998. A review of available systems to investigate transfer of DNA to indigenous soil bacteria. Antonie Leeuwenhoek 73: 15 23.
5. Contreras, A.,, S. Molin,, and J. L. Ramos. 1991. Conditional-suicide containment system for bacteria which mineralize aromatics. Appl. Environ. Microbiol. 57: 1504 1508.
6. Davison, J. 1999. Genetic exchange between bacteria in the environment. Plasmid 42: 73 91.
7. Davison, J. 2002. Genetic tools for pseudomonads, rhizobia, and other gram-negative bacteria. BioTechniques 32: 386 394.
8. Davidson, J . 2002. Towards safer vectors for the environmental release of recombinant bacteria. Environ. Biosafety Res. 1: 9 18.
9. de Lorenzo, V.,, M. Herrero,, J. M. Sánchez,, and K. N. Timmis. 1998. Mini-transposons in microbial ecology and environmental biotechnology. EEMS Microbiol. Ecol. 27: 211 224.
10. Díaz, E.,, M. Munthali,, V. de Lorenzo, and K, N. Timmis. 1994. Universal barrier to lateral spread of specific genes among microorganisms. Mol. Microbiol. 13: 855 861.
11. Díaz, E.,, M. Munthali,, H. Lünsdorf,, J.-V. Höltje,, and K. N, Timmis. 1996. The two-step lysis system of pneumococcal bacteriophage EJ-1 is functional in gram-negative bacteria: triggering of the major pneumococcal autolysin in Escherichia coli. Mol. Microbiol. 19: 667 681.
12. Djordjevic, G. M.,, D. J. O'SulIivan,, S. A. Walker,, M. A. Conkling,, and T. R. Klaenhammer. 1997. A triggered-suicide system designed as a defense against bacteriophages, J. Bacteriol. 179: 6741 6748.
13. Dröge, M.,, A. Pühler,, and W. Selbitschka. 1998. Horizontal gene transfer as a biosafety issue: a natural phenomenon of public concern. J. Biotechnol 64: 75 90.
14. Gerdes, K. 2000. Toxin-antitoxin modules may regulate synthesis of macromolecules during nutritional stress. J. Bacteriol. 182: 561 572.
15. Jensen, L. B.,, J. L. Ramos,, Z. Kaneva,, and S. Molin. 1993. A substrate-dependent biological containment system for Pseudomonas putida based on the Escherichia coli gef gene. Appl. Environ. Microbiol. 59: 3713 3717.
16. Klemm, P.,, L. B. Jensen,, and S. Molin. 1995. A stochastic killing system for biological containment of Escherichia coli. Appl. Environ. Microbiol. 61: 481 486.
17. Kloos, D.-U.,, M. Strätz,, A. Güttler,, R. J. Steffan,, and K. N. Timmis. 1994. Inducible cell lysis system for the study of natural transformation and environmental fate of DNA released by cell death. J. Bacteriol. 176: 7352 7361.
18. Knudsen, S. M.,, and O. H. Karlström. 1991. Development of efficient suicide mechanisms for biological containment of bacteria. Appl Environ. Microbiol 57: 85 92.
19. Knudsen, S.,, P. Saadbye,, L. H. Hansen,, A. Collier,, B. L. Jacobscn,, J. Schlundt,, and O. H. Karlström. 1995. Development and testing of improved suicide functions for biological containment of bacteria. Appl. Environ. Microbiol. 61: 985 991.
20. Kristoffersen, P.,, G. B. Jensen,, K. Gerdes,, and J. Piskur. 2000. Bacterial toxin-antitoxin gene system as containment control in yeast cells. Appl. Environ. Microbiol. 66: 5524 5526.
21. Lorenzo, P.,, S. Alonso,, A. Velasco,, E. Diaz,, J. L. Garcia,, and J. Perera. 2003. Design of catabolic cassettes for styrene biodegradation. Antonie Leeuwenhoek 84: 17 24.
22. Lubitz, W.,, A. Witte,, F. O. Eko,, M. Kamal,, W. Jechlinger,, E. Brand,, J. Marchart,, W. Haidinger,, V. Huter,, D. Felnerova,, N. Stralis-Alves,, S. Lechleitner,, H. Melzer,, M. P. Szostak,, S. Resch,, H. Mader,, B. Kuen,, B. Mayr,, P. Mayrhofer,, R. Gerctsschläger,, A. Haslberger,, and A. Hensel. 1999. Extended recombinant bacterial ghost system. J. Biotechnol. 73: 261 273.
23. Molin, S.,, L. Boc,, L. B. Jensen,, C. S. Kristensen,, M. Givskov,, J. I. Ramos,, and A. K. Bej. 1993. Suicidal genetic elements and their use in biological containment of bacteria. Annu. Rev. Microbiol. 47: 139 166.
24. Molin, S.,, P. Klemm,, L. K. Poulscn,, H. Biehl,, K. Gerdes,, and P. Andersson. 1987. Conditional suicide system for containment of bacteria and plasmids. BioTechnology 5: 1315 1318.
25. Molina, L.,, C. Ramos,, M.-C., Ronchel,, S. Molin,, and J. L. Ramos. 1998. Construction of an efficient biologically contained Pseudomonas putida strain and its survival in outdoor assays. Appl. Environ. Microbiol. 64: 2072 2078.
26. Munthali, M. T.,, K. N. Timmis,, and E. Díaz. 1996. Use of colicin E3 for biological containment of microorganisms. Appl. Environ. Microbiol. 62: 1805 1807.
27. Munthali, M. T.,, K. N. Timmis,, and E. Díaz. 1996. Restricting the dispersal of recombinant DNA: design of a contained biological catalyst. Bio/Technology 14: 189 191.
28. Ramos, J. L.,, P. Andcrsson,, L. B. Jensen,, C. Ramos,, M. C. Ronchel,, E. Díaz,, K. N. Timmis,, and S. Molin. 1995. Suicide microbes on the loose. Bio/Technology 13: 35 37.
29. Recorbet, G.,, C. Robert,, A. Givaudan,, B. Kudla,, P. Normand,, and G. Faurie. 1993. Conditional suicide system of Escherichia coli released into soil that uses the Bacillus subtilis sacB gene. Appl. Environ. Microbiol. 59: 1361 1366.
30. Ronchel, M. C.,, and J. L. Ramos. 2001. Dual system to reinforce biological containment of recombinant bacteria designed for rhizoremcdiation. Appl. Environ. Microbiol. 67: 2649 2656.
31. Ronchel, M. C.,, C. Ramos,, L. B. Jensen,, S. Molin,, and J. L. Ramos. 1995. Construction and behavior of biologically contained bacteria for environmental applications in bioremediation. Appl. Environ. Microbiol. 61: 2990 2994.
32. Schweder, T.,, K. Hofmann,, and M. Hecker. 1995. Escherichia coli K12 relA strains as safe hosts for expression of recombinant DNA. Appl. Microbiol. Biotechnol. 42: 718 723.
33. Schweder, T.,, I. Schmidt,, H. Herrmann,, P. Neubauer,, M. Hecker,, and K. Hofmann. 1992. An expression vector system providing plasmid stability and conditional suicide of plasmid-containing cells. Appl. Microbiol. Biotechnol. 38: 91 93.
34. Sengelov, G.,, and S. J. Sorensen. 1998. Methods for detection of conjugative plasmid transfer in aquatic environments. Curr. MicrobioL. 37: 274 280.
35. Shingler, V.,, and T. Moore. 1994. Sensing of aromatic compounds by the DmpR transcriptional activator of phenol-catabolizing Pseudomonas sp. strain CF600. J. Bacteriol. 176: 1555 1560.
36. Soberón-Chávez, G. 1996. Evaluation of the biological containment system based on the Escherichia coli gef gene in Pseudomonas aeruginosa W51D. Appl. Microbiol. Biotechnol. 46: 549 553.
37. Szafranski, P.,, C. M. Mello,, T. Sano,, C. L. Smith,, D. L. Kaplan,, and C. R. Cantor. 1997. A new approach for containment of microorganisms: dual control of streptavidin expression by antisense RNA and the T7 transcription system. Proc. Natl. Acad. Sci. USA 94: 1059 1063.
38. Tedin, K.,, A. Witte,, G. Reisinger,, W. Lubitz,, and U. Bläsi. 1995. Evaluation of the E. coli ribosomal rrnB P1 promoter and phage-derived lysis genes for the use in a biological containment system: a concept study. J. Biotechnol. 39: 137 148.
39. Torres, B., 2002. Torres, B. 2002, Ph.D. thesis. Universidad Autónoma de Madrid, Madrid, Spain.
40. Torres, B.,, S. Jaenecke,, K. N. Timmis,, J. L. García,, and E. Díaz. 2000. A gene containment strategy based on a restriction- modification system. Environ. Microbiol. 2: 555 563.
41. Vilchez, S.,, L. Molina,, C. Ramos,, and J. L. Ramos. 2000. Proline catabolism by Pseudomonas putida: cloning, characterization, and expression of the put genes in the presence of root exudates. J. Bacteriol. 182: 91 99.
42. Wackett, L. P. 2000. Environmental biotechnology. Trends Biotechnol. 18: 19 21.
43. Young, R.,, I.-N. Wang,, and W. D. Roof. 2000. Phages will out: strategies of host cell lysis. Trends Microbiol. 8: 120 128.

Tables

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Table 1

Lethal functions used in active containment systems

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
Generic image for table
Table 2

Control elements used in active containment systems

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29
Generic image for table
Table 3

Applications of active containment systems

Citation: Torres B, García J, Diaz E. 2004. Plasmids as Tools for Containment, p 589-602. In Funnell B, Phillips G (ed), Plasmid Biology. ASM Press, Washington, DC. doi: 10.1128/9781555817732.ch29

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