Description:
<jats:p><jats:bold>Aim.</jats:bold>The nascent field of bio-geoengineering stands to benefit from synthetic biologists’ efforts to standardise, and in so doing democratise, biomolecular research methods.<jats:italic>Roseobacter</jats:italic>clade bacteria comprise 15–20% of oceanic bacterio-plankton communities, making them a prime candidate for establishment of synthetic biology chassis for bio-geoengineering activities such as bioremediation of oceanic waste plastic. Developments such as the increasing affordability of DNA synthesis and laboratory automation continue to foster the establishment of a global ‘do-it-yourself’ research community alongside the more traditional arenas of academe and industry. As a collaborative group of citizen, student and professional scientists we sought to test the following hypotheses: (i) that an incubator capable of cultivating bacterial cells can be constructed entirely from non-laboratory items, (ii) that marine bacteria from the<jats:italic>Roseobacter</jats:italic>clade can be established as a genetically tractable synthetic biology chassis using plasmids conforming to the BioBrick<jats:sup>TM</jats:sup>standard and finally, (iii) that identifying and subcloning genes from a<jats:italic>Roseobacter</jats:italic>clade species can readily by achieved by citizen scientists using open source cloning and bioinformatic tools.</jats:p><jats:p><jats:bold>Method.</jats:bold>We cultivated three<jats:italic>Roseobacter</jats:italic>species,<jats:italic>Roseobacter denitrificans</jats:italic>,<jats:italic>Oceanobulbus indolifex</jats:italic>and<jats:italic>Dinoroseobacter shibae</jats:italic>. For each species we measured chloramphenicol sensitivity, viability over 11 weeks of glycerol-based cryopreservation and tested the effectiveness of a series of electroporation and heat shock protocols for transformation using a variety of plasmid types. We also attempted construction of an incubator-shaker device using only publicly available components. Finally, a subgroup comprising citizen scientists designed and attempted a procedure for isolating the cold resistance<jats:italic>anf1</jats:italic>gene from<jats:italic>Oceanobulbus indolifex</jats:italic>cells and subcloning it into a BioBrick<jats:sup>TM</jats:sup>formatted plasmid.</jats:p><jats:p><jats:bold>Results.</jats:bold>All species were stable over 11 weeks of glycerol cryopreservation, sensitive to 17 µg/mL chloramphenicol and resistant to transformation using the conditions and plasmids tested. An incubator-shaker device, ‘UCLHack-12’ was assembled and used to cultivate sufficient quantity of<jats:italic>Oceanobulbus indolifex</jats:italic>cells to enable isolation of the<jats:italic>anf1</jats:italic>gene and its subcloning into a plasmid to generate the BioBrick<jats:sup>TM</jats:sup>BBa_K729016.</jats:p><jats:p><jats:bold>Conclusion.</jats:bold>The process of ‘de-skilling’ biomolecular techniques, particularly for relatively under-investigated organisms, is still on-going. However, our successful cell growth and DNA manipulation experiments serve to indicate the types of capabilities that are now available to citizen scientists. Science democratised in this way can make a positive contribution to the debate around the use of bio-geoengineering to address oceanic pollution or climate change.</jats:p>