Accurate multiplex gene synthesis from programmable DNA microchips

Testing the many hypotheses from genomics and systems biology experiments demands accurate and cost-effective gene and genome synthesis. Here we describe a microchip-based technology for multiplex gene synthesis. Pools of thousands of 'construction' oligonucleotides and tagged complementary 'selection' oligonucleotides are synthesized on photo-programmable microfluidic chips, released, amplified and selected by hybridization to reduce synthesis errors ninefold. A one-step polymerase assembly multiplexing reaction assembles these into multiple genes. This technology enabled us to synthesize all 21 genes that encode the proteins of the Escherichia coli 30S ribosomal subunit, and to optimize their translation efficiency in vitro through alteration of codon bias. This is a significant step towards the synthesis of ribosomes in vitro and should have utility for synthetic biology in general.     

Active foundering of a continental arc root beneath the southern Sierra Nevada in California

Seismic data provide images of crust-mantle interactions during ongoing removal of the dense batholithic root beneath the southern Sierra Nevada mountains in California. The removal appears to have initiated between 10 and 3 Myr ago with a Rayleigh-Taylor-type instability, but with a pronounced asymmetric flow into a mantle downwelling (drip) beneath the adjacent Great Valley. A nearly horizontal shear zone accommodated the detachment of the ultramafic root from its granitoid batholith. With continuing flow into the mantle drip, viscous drag at the base of the remaining approximately 35-km-thick crust has thickened the crust by approximately 7 km in a narrow welt beneath the western flank of the range. Adjacent to the welt and at the top of the drip, a V-shaped cone of crust is being dragged down tens of kilometres into the core of the mantle drip, causing the disappearance of the Moho in the seismic images. Viscous coupling between the crust and mantle is therefore apparently driving present-day surface subsidence.     

Genome sequence of the Brown Norway rat yields insights into mammalian evolution

The laboratory rat (Rattus norvegicus) is an indispensable tool in experimental medicine and drug development, having made inestimable contributions to human health. We report here the genome sequence of the Brown Norway (BN) rat strain. The sequence represents a high-quality 'draft' covering over 90% of the genome. The BN rat sequence is the third complete mammalian genome to be deciphered, and three-way comparisons with the human and mouse genomes resolve details of mammalian evolution. This first comprehensive analysis includes genes and proteins and their relation to human disease, repeated sequences, comparative genome-wide studies of mammalian orthologous chromosomal regions and rearrangement breakpoints, reconstruction of ancestral karyotypes and the events leading to existing species, rates of variation, and lineage-specific and lineage-independent evolutionary events such as expansion of gene families, orthology relations and protein evolution.     

Derivation of embryonic germ cells and male gametes from embryonic stem cells

Egg and sperm cells (gametes) of the mouse are derived from a founder population of primordial germ cells that are set aside early in embryogenesis. Primordial germ cells arise from the proximal epiblast, a region of the early mouse embryo that also contributes to the first blood lineages of the embryonic yolk sac. Embryonic stem cells differentiate in vitro into cystic structures called embryoid bodies consisting of tissue lineages typical of the early mouse embryo. Because embryoid bodies sustain blood development, we reasoned that they might also support primordial germ cell formation. Here we isolate primordial germ cells from embryoid bodies, and derive continuously growing lines of embryonic germ cells. Embryonic germ cells show erasure of the methylation markers (imprints) of the Igf2r and H19 genes, a property characteristic of the germ lineage. We show that embryoid bodies support maturation of the primordial germ cells into haploid male gametes, which when injected into oocytes restore the somatic diploid chromosome complement and develop into blastocysts. Our ability to derive germ cells from embryonic stem cells provides an accessible in vitro model system for studies of germline epigenetic modification and mammalian gametogenesis.     

Convergence across biomes to a common rain-use efficiency

Water availability limits plant growth and production in almost all terrestrial ecosystems. However, biomes differ substantially in sensitivity of aboveground net primary production (ANPP) to between-year variation in precipitation. Average rain-use efficiency (RUE; ANPP/precipitation) also varies between biomes, supposedly because of differences in vegetation structure and/or biogeochemical constraints. Here we show that RUE decreases across biomes as mean annual precipitation increases. However, during the driest years at each site, there is convergence to a common maximum RUE (RUE(max)) that is typical of arid ecosystems. RUE(max) was also identified by experimentally altering the degree of limitation by water and other resources. Thus, in years when water is most limiting, deserts, grasslands and forests all exhibit the same rate of biomass production per unit rainfall, despite differences in physiognomy and site-level RUE. Global climate models predict increased between-year variability in precipitation, more frequent extreme drought events, and changes in temperature. Forecasts of future ecosystem behaviour should take into account this convergent feature of terrestrial biomes.     

Electrolytic reduction of mixed solid oxides in molten salts for energy efficient production of the TiNi alloy

Fast and energy efficient technologies for material production are essential for the sustainable develop- ment of human society. The TiNi alloy is an important intelligent material[1], having high abrasive resistance, superior elasticity and good shape me…     

Federal Republic of Germany The Annual Report for 1987 of the Deutsche Forschungsgemeinschaft (German Research Association)

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Federal Republic of Germany The Annual Report for 1987 of the Deutsche Forschungsgemeinschaft (German Research Association)