Doing synthetic biology with photosynthetic microorganisms

The use of photosynthetic microbes as synthetic biology hosts for the sustainable production of commodity chemicals and even fuels has received increasing attention over the last decade. The number of studies published, tools implemented, and resources made available for microalgae have increased beyond expectations during the last few years. However, the tools available for genetic engineering in these organisms still lag those available for the more commonly used heterotrophic host organisms. In this mini-review, we provide an overview of the photosynthetic microbes most commonly used in synthetic biology studies, namely cyanobacteria, chlorophytes, eustigmatophytes and diatoms. We provide basic information on the techniques and tools available for each model group of organisms, we outline the state-of-the-art, and we list the synthetic biology tools that have been successfully used. We specifically focus on the latest CRISPR developments, as we believe that precision editing and advanced genetic engineering tools will be pivotal to the advancement of the field. Finally, we discuss the relative strengths and weaknesses of each group of organisms and examine the challenges that need to be overcome to achieve their synthetic biology potential.Peer reviewe

Investigations of the Natural Fission Reactor Program. Progress report, October 1976--September 1977

During Fiscal Year 1977, efforts of the Natural Fission Reactor Program have been directed toward (1) further measurements and predictions of reactor product migration at Oklo, (2) uraninite solubility studies, (3) radiogenic equilibria studies of natural Pu, Ra, and Th in U ores, (4) search for evidence of other natural fission reactors, and (5) supportive analytical measurements and procedures development

Search for other natural fission reactors

Precambrian uranium ores have been surveyed for evidence of other natural fission reactors. The requirements for formation of a natural reactor direct investigations to uranium deposits with large, high-grade ore zones. Massive zones with volumes approximately greater than 1 m/sup 3/ and concentrations approximately greater than 20 percent uranium are likely places for a fossil reactor if they are approximately greater than 0.6 b.a. old and if they contained sufficient water but lacked neutron-absorbing impurities. While uranium deposits of northern Canada and northern Australia have received most attention, ore samples have been obtained from the following worldwide locations: the Shinkolobwe and Katanga regions of Zaire; Southwest Africa; Rio Grande do Norte, Brazil; the Jabiluka, Nabarlek, Koongarra, Ranger, and El Sharana ore bodies of the Northern Territory, Australia; the Beaverlodge, Maurice Bay, Key Lake, Cluff Lake, and Rabbit Lake ore bodies and the Great Bear Lake region, Canada. The ore samples were tested for isotopic variations in uranium, neodymium, samarium, and ruthenium which would indicate natural fission. Isotopic anomalies were not detected. Criticality was not achieved in these deposits because they did not have sufficient /sup 235/U content (a function of age and total uranium content) and/or because they had significant impurities and insufficient moderation. A uranium mill monitoring technique has been considered where the ''yellowcake'' output from appropriate mills would be monitored for isotopic alterations indicative of the exhumation and processing of a natural reactor

Genetic engineering of the multicellular green alga Volvox: a modified and multiplied bacterial antibiotic resistance gene as a dominant selectable marker

Hallmann A, Rappel A. Genetic engineering of the multicellular green alga Volvox: a modified and multiplied bacterial antibiotic resistance gene as a dominant selectable marker. Plant J. 1999;17(1):99-109.The green alga Volvox represents the simplest multicellular organism: Volvox is composed of only two cell types, somatic and reproductive. Volvox, therefore, is an attractive model system for studying various aspects of multicellularity. With the biolistic nuclear transformation of Volvox carteri, the powerful molecular genetic manipulation of this organism has been established, but applications have been restricted to an auxotrophic mutant serving as the DNA recipient. Therefore, a dominant selectable marker working in all strains and mutants of this organism is required. Among several gene constructs tested, the most advantageous results were obtained with a chimeric gene composed of the coding sequence of the bacterial ble gene, conferring resistance to the antibiotic zeocin, modified with insertions of two endogenous introns from the Volvox arylsulfatase gene and fused to 5' and 3' untranslated regions from the Volvox beta 2-tubulin gene. In the most suitable plasmid used, the gene dosage was increased 16-fold by a technique that allows exponential multiplication of a DNA fragment. Co-transformation of this plasmid and a non-selectable plasmid allowed the identification of zeocin resistant transformants with nuclear integration of both selectable and non-selectable plasmids. Stable expression of the ble gene and of genes from several non-selectable plasmids is demonstrated. The modified ble gene provides the first dominant marker for transformation of both wild-type and mutant strains of Volvox