scholarly journals Rapid Golden Gate assembly of exons from genomic DNA for protein expression in Escherichia coli and Pichia pastoris

BioTechniques ◽  
2021 ◽  
Author(s):  
Junhao Cheng ◽  
Mingkun Wu ◽  
Ren Zhong ◽  
Dayong Si ◽  
Geng Meng ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pichia Pastoris ◽  
Genomic Dna ◽  
Low Cost ◽  
Cost Effective ◽  
Single Step ◽  
Golden Gate ◽  
Cloning Efficiency ◽  
Expression In Escherichia Coli ◽  
Expression Plasmids

The development of a quick, single-step cloning system for generation of multiexon gene expression constructs is presented. The system allows efficient and cost-effective assembly of multiple exons of interest genes into different expression plasmids in both Escherichia coli and Pichia pastoris. The high cloning efficiency and low cost of the system make it ideal for a novel workflow for the assembly of intron-bearing genes for expression in two different expression hosts.

scholarly journals Strain-enabled ultra-broadband Ge-based photonic devices for low-cost integration in PICs

2020 ◽  
Author(s):  
Yiding Lin ◽  
Danhao Ma ◽  
Rui-Tao Wen ◽  
Kwang Hong Lee ◽  
Govindo Syaranamual ◽  
...  
Keyword(s):  
Integrated Circuits ◽  
Communication Networks ◽  
High Efficiency ◽  
Low Cost ◽  
Data Communication ◽  
Cost Effective ◽  
Photonic Devices ◽  
Single Step ◽  
High Data ◽  
Data Capacity

Abstract Photonic-integrated circuits (PICs) have become one of the most promising solutions to the burgeoning global data communication and are being envisioned to have revolutionary impact in many other emerging fields. This outlook requires future PICs to be significantly more broadband and cost-effective. The current germanium (Ge)-based active photonic devices in PICs are thus facing a new bandwidth-cost trade-off. Here, we demonstrate ultra-broadband, high-efficiency Ge photodetectors up to 1,630 nm operation wavelength and Ge0.99Si0.01 electro-absorption (EA) modulator arrays with an operating range of ~100 nm from 1,525 to 1,620 nm, using a CMOS-compatible recess-type silicon nitride (SiNx) stressor. The broadband operation could facilitate a wide (>100 nm) window for low-cost Ge modulator-detector co-integration, requiring only a single step of Ge epitaxy and two different SiNx depositions. The broad modulation and co-integration coverage can be entirely shifted to shorter (~1,300 nm) and longer (>1,700 nm) wavelengths with small amounts of Si or tin (Sn) alloying. This proof-of-concept work provides a pathway for PICs towards future low-cost and high-data-capacity communication networks, immediately accessible by designers through foundries.

Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris

2011 ◽  
Vol 111 (5) ◽  
pp. 528-536 ◽  
Author(s):  
Thayat Sriyapai ◽  
Peechapack Somyoonsap ◽  
Kenji Matsui ◽  
Fusako Kawai ◽  
Kosum Chansiri
Keyword(s):  
Escherichia Coli ◽  
Pichia Pastoris ◽  
Thermostable Xylanase ◽  
Expression In Escherichia Coli

Cost-Effective Single Step Cofiring Process for Manufacturing Solid Oxide Fuel Cells Using HSC™ Anode

2010 ◽  
Vol 7 (2) ◽  
Author(s):  
Kyung Joong Yoon ◽  
Guosheng Ye ◽  
Srikanth Gopalan ◽  
Uday B. Pal
Keyword(s):  
Active Layer ◽  
Low Cost ◽  
Cost Effective ◽  
High Volume ◽  
Single Step ◽  
Solid Oxide ◽  
Fine Grained ◽  
High Volume Production ◽  
Volume Production ◽  
Anode Substrate

The anode-supported planar solid oxide fuel cell (SOFC) was fabricated by a cost-effective single step cofiring process using high shear compaction (HSC)™ anode substrate. The HSC™ process is a novel ceramic tape fabrication technique, which offers advantages in low-cost and high-volume production of the anode substrates over the conventional tape forming processes. The cell was comprised of a porous HSC™ Ni+8 mol % yttria-stabilized zirconia (YSZ) anode substrate, a porous Ni+YSZ anode barrier layer, a porous and fine-grained Ni+YSZ anode active layer, a dense YSZ electrolyte, a porous and fine-grained Ca-doped LaMnO3(LCM)+YSZ composite cathode active layer, and a porous LCM cathode current collector layer. The fabrication process involved wet powder spraying of the anode barrier layer over the HSC™ anode substrate followed by screen-printing of the remaining component layers. The cell was then cofired at 1340°C for 2 h. The microstructure and the open circuit voltage of the cell confirmed that the cell was crack-free and leak-tight. The cofired cell showed a stable and acceptable electrochemical performance at 800°C under humidified hydrogen (3–60% H2O) as fuel and air as oxidant. The anode active layer with finer and less porous microstructure increased the triple phase boundary length and improved cell performance under conditions that simulated higher fuel utilization. The material system and fabrication process presented in this work offers great advantage in low-cost and high-volume production of SOFCs, and it can be the basis for scale-up and successful commercialization of the SOFC technology.

scholarly journals Novel biotechnological approaches for monitoring and immunization against resistant to antibiotics Escherichia coli and other pathogenic bacteria

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
José E. Belizário ◽  
Marcelo P. Sircili
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Pathogenic Bacteria ◽  
Low Cost ◽  
Methyl Esters ◽  
Cost Effective ◽  
Antibiotic Use ◽  
Immunological Methods ◽  
Bacterial Strains ◽  
Surveillance Programs

Abstract The application of next-generation molecular, biochemical and immunological methods for developing new vaccines, antimicrobial compounds, probiotics and prebiotics for zoonotic infection control has been fundamental to the understanding and preservation of the symbiotic relationship between animals and humans. With increasing rates of antibiotic use, resistant bacterial infections have become more difficult to diagnose, treat, and eradicate, thereby elevating the importance of surveillance and prevention programs. Effective surveillance relies on the availability of rapid, cost-effective methods to monitor pathogenic bacterial isolates. In this opinion article, we summarize the results of some research program initiatives for the improvement of live vaccines against avian enterotoxigenic Escherichia coli using virulence factor gene deletion and engineered vaccine vectors based on probiotics. We also describe methods for the detection of pathogenic bacterial strains in eco-environmental headspace and aerosols, as well as samples of animal and human breath, based on the composition of volatile organic compounds and fatty acid methyl esters. We explain how the introduction of these low-cost biotechnologies and protocols will provide the opportunity to enhance co-operation between networks of resistance surveillance programs and integrated routine workflows of veterinary and clinical public health microbiology laboratories.

Secretory expression in Escherichia coli and single-step purification of a heat-stable alkaline protease

2003 ◽  
Vol 28 (1) ◽  
pp. 63-68 ◽  
Author(s):  
Zhibiao Fu ◽  
Suhali Bt Ab Hamid ◽  
Che Nyonya A Razak ◽  
Mahiran Basri ◽  
Abu Bakar Salleh ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Alkaline Protease ◽  
Single Step ◽  
Secretory Expression ◽  
Expression In Escherichia Coli ◽  
Heat Stable ◽  
Single Step Purification

Fabrication of Cost-Effective Polymer-Based Nanofluidic Device for Single Molecular Analysis

2010 ◽  
Author(s):  
Jiahao Wu ◽  
Rattikan Chantiwas ◽  
Steven A. Soper ◽  
Sunggook Park
Keyword(s):  
Focused Ion Beam ◽  
Low Cost ◽  
Ion Beam ◽  
Cost Effective ◽  
Single Step ◽  
Molecular Sensor ◽  
Sensor Structure ◽  
Nanofluidic Devices ◽  
Nanofluidic Device ◽  
Hierarchical Multiscale

A nanochannel based single molecular sensor was fabricated in poly(methyl methacrylate) substrate by a single step imprinting process with a polymer stamp. The sensor structure consists of hierarchical multiscale patterns of an array of 90 nm (depth) × 190 nm (width) × 80 μm (length) nanochannels and microfluidic networks. In contrast to nanofluidic devices fabricated by high-end nanofabrication tools such as focused ion beam milling and electron beam lithography, this direct imprint process is more desirable for efficient and low cost fabrication. Moreover, due to the reduction of stress generated during imprinting, this polymer stamp imprinting process can also reduce the deformation (warping) in molded substrates, and prevent the damage of expensive nanostructured stamp as well.

scholarly journals One-Step Production of Immobilized α-Amylase in Recombinant Escherichia coli

2009 ◽  
Vol 75 (7) ◽  
pp. 2012-2016 ◽  
Author(s):  
Indira A. Rasiah ◽  
Bernd H. A. Rehm
Keyword(s):  
Escherichia Coli ◽  
Structural Integrity ◽  
Amylase Activity ◽  
Signal Sequence ◽  
Cost Effective ◽  
Single Step ◽  
Enzyme Analysis ◽  
Fusion Partner ◽  
One Step ◽  
The Stability

ABSTRACT Industrial enzymes are often immobilized via chemical cross-linking onto solid supports to enhance stability and facilitate repeated use in bioreactors. For starch-degrading enzymes, immobilization usually places constraints on enzymatic conversion due to the limited diffusion of the macromolecular substrate through available supports. This study describes the one-step immobilization of a highly thermostable α-amylase (BLA) from Bacillus licheniformis and its functional display on the surface of polyester beads inside engineered Escherichia coli. An optimized BLA variant (Termamyl) was N-terminally fused to the polyester granule-forming enzyme PhaC of Cupriavidus necator. The fusion protein lacking the signal sequence mediated formation of stable polyester beads exhibiting α-amylase activity. The α-amylase beads were assessed with respect to α-amylase activity, which was demonstrated qualitatively and quantitatively. The immobilized α-amylase showed Michaelis-Menten enzyme kinetics exerting a V max of about 506 mU/mg of bead protein with a Km of about 5 μM, consistent with that of free α-amylase. The stability of the enzyme at 85°C and the capacity for repeated usage in a starch liquefaction process were also demonstrated. In addition, structural integrity and functionality of the beads at extremes of pH and temperature, demonstrating their suitability for industrial use, were confirmed by electron microscopy and protein/enzyme analysis. This study proposes a novel, cost-effective method for the production of immobilized α-amylase in a single step by using the polyester granules forming protein PhaC as a fusion partner in engineered E. coli.

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