Simplified transformation of ostreococcus tauri using polyethylene glycol

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. Ostreococcustauri is an easily cultured representative of unicellular algae (class Mamiellophyceae) that abound in oceans worldwide. Eight complete 13–22 Mb genomes of phylogenetically divergent species within this class are available, and their DNA sequences are nearly always present in metagenomic data produced from marine samples. Here we describe a simplified and robust transformation protocol for the smallest of these algae (O. tauri). Polyethylene glycol (PEG) treatment was much more effcient than the previously described electroporation protocol. Short (2 min or less) incubation times in PEG gave >104 transformants per microgram DNA. The time of cell recovery after transformation could be reduced to a few hours, permitting the experiment to be done in a day rather than overnight as used in previous protocols. DNA was randomly inserted in the O. tauri genome. In our hands PEG was 20–40-fold more effcient than electroporation for the transformation of O. tauri, and this improvement will facilitate mutagenesis of all of the dispensable genes present in the tiny O. tauri genome

Resin infusion of layered metal/composite hybrid and resulting metal/composite hybrid laminate

A method of fabricating a metal/composite hybrid laminate is provided. One or more layered arrangements are stacked on a solid base to form a layered structure. Each layered arrangement is defined by a fibrous material and a perforated metal sheet. A resin in its liquid state is introduced along a portion of the layered structure while a differential pressure is applied across the laminate structure until the resin permeates the fibrous material of each layered arrangement and fills perforations in each perforated metal sheet. The resin is cured thereby yielding a metal/composite hybrid laminate

Elevated Temperature, Notched Compression Performance of Out of Autoclave Processed Composites

Curved honeycomb sandwich panels composed of carbon fiber reinforced toughened-epoxy polymer facesheets are being evaluated for potential use as payload fairing components on the NASA heavy-lift space launch system (HL-SLS). These proposed composite sandwich panels provide the most efficient aerospace launch structures, and offer mass and thermal advantages when compared with existing metallic payload fairing structures. NASA and industry are investigating recently developed carbon fiber epoxy prepreg systems which can be fabricated using out-of autoclave (OOA) processes. Specifically, OOA processes using vacuum pressure in an oven and thereby significantly reducing the cost associated with manufacturing large (up to 10 m diameter) composite structures when compared with autoclave. One of these OOA composite material systems, CYCOM(R) 5320-1, was selected for manufacture of a 1/16th scale barrel portion of the payload fairing; such that, the system could be compared with the well-characterized prepreg system, CYCOM(R) 977-3, typically processed in an autoclave. Notched compression coupons for each material were obtained from the minimum-gauge flat laminate [60/-60/0]S witness panels produced in this manufacturing study. The coupons were also conditioned to an effective moisture equilibrium point and tested according to ASTM D6484M-09 at temperatures ranging from 25 C up to 177 C. The results of this elevated temperature mechanical characterization study demonstrate that, for thin coupons, the OHC strength of the OOA laminate was equivalent to the flight certified autoclave processed composite laminates; the limitations on the elevated temperature range are hot-wet conditions up to 163 C and are only within the margins of testing error. At 25 C, both the wet and dry OOA material coupons demonstrated greater OHC failure strengths than the autoclave processed material laminates. These results indicate a substantial improvement in OOA material development and processing since previous studies have consistently reported OOA material strengths on par or below those of autoclave processed composite laminates

Navy design spending in an era of acquisition reform

Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Ocean Engineering, 1999.Includes bibliographical references (leaf 153).by Jennifer S. Grimsley.S.M

Elevated Temperature, Residual Compressive Strength of Impact-Damaged Sandwich Structure Manufactured Out-of-Autoclave

Several 1/16th-scale curved sandwich composite panel sections of a 10 m diameter barrel were fabricated to demonstrate the manufacturability of large-scale curved sections using minimum gauge, [+60/-60/0]s, toughened epoxy composite facesheets co-cured with low density (50 kilograms per cubic meters) aluminum honeycomb core. One of these panels was fabricated out of autoclave (OoA) by the vacuum bag oven (VBO) process using Cycom(Registered Trademark) T40-800b/5320-1 prepreg system while another panel with the same lay-up and dimensions was fabricated using the autoclave-cure, toughened epoxy prepreg system Cycom(Registered Trademark) IM7/977-3. The resulting 2.44 m x 2 m curved panels were investigated by non-destructive evaluation (NDE) at NASA Langley Research Center (NASA LaRC) to determine initial fabrication quality and then cut into smaller coupons for elevated temperature wet (ETW) mechanical property characterization. Mechanical property characterization of the sandwich coupons was conducted including edge-wise compression (EWC), and compression-after-impact (CAI) at conditions ranging from 25 C/dry to 150 C/wet. The details and results of this characterization effort are presented in this paper

Paper Session II-A - Polyimide Foam Insulation Materials for Aerospace Vehicles and Spaceport Applications

Advancements in high temperature materials by NASA have led to the development of polyimide foam systems with very attractive properties. The properties generated demonstrate the suitability of these materials for use as insulation for cryogenic fuel tanks on next generation vehicles, commercial and military ships, and potentially commercial aircraft. The significance of structural polyimide foams can be realized with a reduction in the overall weight of a launch vehicle. Due to a polyimide\u27 s high operating temperature ( \u27 \u27 260°C) structural polyimide foams can potentially reduce the amount of Thermal Protection System (TPS) and TPS integration structure that is required on launch vehicles. The lowtemperature elasticity of other polyimide foams is an enabling feature for many new cryogenic applications. These high performance materials also have properties that fulfill the demanding upcoming needs in ground support equipment for a Spaceport Technology Center. In a research study performed by Kennedy Space Center (KSC) and Langley Research Center (LaRC), polyimide foams were investigated for their physical, mechanical, thermal, and flammability properties. Variations in chemical structure, cell surface area, cell content and density on the resultant physical properties of the foams were studied. Data generated from this research revealed vital information involving foam technology and the interplay of factors such as foam density, open-closed cell content, surface area, and cell structure on the overall performance of the material. By controlling these parameters, new thermal insulation systems based on polyimide foam materials can be designed to meet demanding applications for spaceports and space vehicles

How and why DNA barcodes underestimate the diversity of microbial eukaryotes

Background: Because many picoplanktonic eukaryotic species cannot currently be maintained in culture, direct sequencing of PCR-amplified 18S ribosomal gene DNA fragments from filtered sea-water has been successfully used to investigate the astounding diversity of these organisms. The recognition of many novel planktonic organisms is thus based solely on their 18S rDNA sequence. However, a species delimited by its 18S rDNA sequence might contain many cryptic species, which are highly differentiated in their protein coding sequences. Principal Findings: Here, we investigate the issue of species identification from one gene to the whole genome sequence. Using 52 whole genome DNA sequences, we estimated the global genetic divergence in protein coding genes between organisms from different lineages and compared this to their ribosomal gene sequence divergences. We show that this relationship between proteome divergence and 18S divergence is lineage dependant. Unicellular lineages have especially low 18S divergences relative to their protein sequence divergences, suggesting that 18S ribosomal genes are too conservative to assess planktonic eukaryotic diversity. We provide an explanation for this lineage dependency, which suggests that most species with large effective population sizes will show far less divergence in 18S than protein coding sequences. Conclusions: There is therefore a trade-off between using genes that are easy to amplify in all species, but which by their nature are highly conserved and underestimate the true number of species, and using genes that give a better description of the number of species, but which are more difficult to amplify. We have shown that this trade-off differs between unicellular and multicellular organisms as a likely consequence of differences in effective population sizes. We anticipate that biodiversity of microbial eukaryotic species is underestimated and that numerous ''cryptic species'' will become discernable with the future acquisition of genomic and metagenomic sequences