Optimizing the process of genetic material transmission using bacterial conjugation (Konyugatsia-EFR) - 03.14.18

Overview | Description | Applications | Operations | Results | Publications | Imagery

ISS Science for Everyone

Science Objectives for Everyone
The Konyugatsiya biotechnology experiment is devoted to the development of methods for constructing new recombinant strains producing biologically active substances (BAS) via the transmission of plasmid and chromosome DNA using bacterial conjugation. This experiment calls for the combination, in a reactor vessel, of liquid donor and recipient strain cultures under space flight conditions with subsequent genetic analysis of obtained cultures and selection of stable hybrids on the ground. The research enables scientists to develop methods of obtaining hybrid strains producing BAS via the transmission of plasmid and chromosome DNA using bacterial conjugation.
Science Results for Everyone
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The following content was provided by Yu. P. Zerov, and is maintained in a database by the ISS Program Science Office.
Experiment Details

OpNom:

Principal Investigator(s)
Yu. P. Zerov, Biopreparat, Russia

Co-Investigator(s)/Collaborator(s)
Information Pending

Developer(s)
Information Pending

Sponsoring Space Agency
Russian Federal Space Agency (Roscosmos)

Sponsoring Organization
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Research Benefits
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ISS Expedition Duration
April 2003 - October 2003; April 2004 - April 2005; October 2005 - October 2008

Expeditions Assigned
7,9,10,12,13,14,15,16,17

Previous Missions
Information Pending

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Experiment Description

Research Overview
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Description
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Applications

Space Applications
Data obtained on the influence of orbital flight conditions on the effectiveness of transferring chromosome and plasmid DNA using bacterial conjunction may be used to create process lines to produce recombinant proteins under microgravity conditions.

Earth Applications
The developed method of obtaining hybrid strains producing BAS through the transfer of chromosome or plasmid DNA using bacterial conjugation under space flight conditions and subsequent ground selection may be used to obtain producers of proteins that are valuable for health care.

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Operations

Operational Requirements and Protocols
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Decadal Survey Recommendations

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Results/More Information

Recombinant donor and recipient strains are engineered on the ground for each experiment session. As a result of implementing two Konyugatsiya experiments during ISS Expeditions 23/24 and 27/28, two recombinant plasmids that encode the end human Cu, Zn superoxide dismutase (SOD) protein were successfully transferred. From the biomaterial obtained during conjugative hybridization in the experiment, four potential variant hybrid clones of SOD-producing E. coli were identified: two groups depending on the recipient strain and plasmid and two hybrid alternatives each in each group: type I hybrids, which received only the recombinant plasmid as a result of mobilization, and type II hybrids, which also received the F’ conjugative plasmid. The mean productivity of the obtained clones of the two variants of strains producing SOD was 170–185 μg/ml of culture, which is sufficiently high for primary clones. In both cases the productivity of obtained lines of hybrid strains is sufficiently high, as to allow them to be used as original material for the selection of high-productivity producer strain variants.

Another object of study that is of great practical interest is the human epidermal growth factor (EGF). EGF is a unique growth factor that ensures monolayer skin growth, i.e., wound healing without the formation of scar tissue. This factor has ensured its wide use in treating burns, for healing post-surgical sutured incisions, for treating ischemic ulcers, bed sores, and freezing injuries, for preventing and treating radiation dermatitis, as well as for cosmetic surgery. Obtaining hybrid producers of EGF in the Konyugatsiya experiment met with serious difficulties. Owing to the high toxicity of the end product for bacteria cells, the obtained hybrids gradually lost the ability to synthesize EGF. Finally, as a result of the Konyugatsiya experiment implemented during ISS Expedition 39/40, two stable and pure lines of a strain producing EGF—E. coli BL21(DE3)/F′;pEThEGFоriT—were obtained by combining the preliminary selection in the experiment with the subsequent ground-based selection. Both obtained lines of strains producing EGF exhibited high genetic stability and reproducibility with respect to end product output.

Obtained variants of the strain E. coli BL21(DE3)/F′;pEThEGFоriT are of interest as promising sources for the production of the EGF-producing strain. Results confirmed that the method of mobilizing recombinant plasmids in bacterial conjugation in the experiment allows bacterial producer strains with a levels of end product biosynthesis high enough for practical use to be obtained.

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Related Websites
Energia - Science Research on the ISS Russian Segment

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Imagery

image Thermal insulating container 'Biokont-T', Recomb-K equipment, thermostatically controlled container 'Akvarius'.
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