Design and construction of an in-plant activation cassette for transgene expression and recombinant protein production in plants

Virus-based transgene expression systems have become particularly valuable for recombinant protein production in plants. The dual-module in-plant activation (INPACT) expression platform consists of a uniquely designed split-gene cassette incorporating the cis replication elements of Tobacco yellow dwarf geminivirus (TYDV) and an ethanol-inducible activation cassette encoding the TYDV Rep and RepA replication-associated proteins. The INPACT system is essentially tailored for recombinant protein production in stably transformed plants and provides both inducible and high-level transient transgene expression with the potential to be adapted to diverse crop species. The construction of a novel split-gene cassette, the inducible nature of the system and the ability to amplify transgene expression via rolling-circle replication differentiates this system from other DNA- and RNA-based virus vector systems used for stable or transient recombinant protein production in plants. Here we provide a detailed protocol describing the design and construction of a split-gene INPACT cassette, and we highlight factors that may influence optimal activation and amplification of gene expression in transgenic plants. By using Nicotiana tabacum, the protocol takes 6-9 months to complete, and recombinant proteins expressed using INPACT can accumulate to up to 10% of the leaf total soluble protein.

Expression of a bacterial xylanase in Trichoderma reesei under the egl2 and cbh2 glycosyl hydrolase gene promoters

Expression vectors were constructed for Trichoderma reesei using the promoters, secretion signals and the modular structure of the efficiently expressed and secreted cellulase enzymes EGL2 (Cel5A) and CBH2 (Cel6A) as a prelude to establishing a platform where a gene of interest can be expressed under several promoters simultaneously. The designs featured (i) EGL2sigpro (egl2 promoter and secretion signal), (ii) EGL2cbmlin (egl2 promoter, secretion signal, EGL2 cellulose binding module and linker), (iii) CBH2sigpro (cbh2 promoter and secretion signal) and (iv) CBH2cbmlin (cbh2 promoter, secretion signal, CBH2 cellulose binding module and linker). Recombinant vectors were introduced individually into the high protein-secreting T. reesei RUT-C30 strain to generate single-promoter transformants expressing the Dictyoglomus thermophilum xynB gene that encodes a thermophilic xylanase enzyme (XynB). Ten transformants producing XynB representing each of the four different types of vectors were selected for further testing and the highest XynB production was achieved from a transformant containing 1–2 copies of the EGL2cbmlin vector. Best xylanase producers did not show any particular pattern in terms of the number of gene copies and their mode of integration into the chromosomal DNA. Transformants generated with the cbmlin-type vectors produced multiple forms of XynB which were decorated with various N- and O-glycans. One of the O-glycans was identified as hexuronic acid, whose presence had not been observed previously in the glycosylation patterns of T. reesei.

A platform - independent product library for BIM

There is considerable interest internationally in developing product libraries to support the use of BIM. Product library initiatives are driven by national bodies, manufacturers and private companies who see their potential. A major issue with the production and distribution of product information for BIM is that separate library objects need to be produced for all of the different software systems that are going to use the library. This increases the cost of populating product libraries and also increases the difficulty in maintaining consistency between the representations for the different software over time. This paper describes a project which uses “software transformation” technology from the field of software engineering to support the definition of a single generic representation of a product which can then be automatically converted to the format required by receiving software. The paper covers the current state of implementation of the product library, the technology underlying the transformations for the currently supported software and the business model for creating a national library in Australia. This is placed within the context of other current product library systems to highlight the differences. The responsibilities of the various actors involved in supporting the product library are also discussed.

Development of an inter-vehicle communications & positioning platform for transport safety applications

This project is a breakthrough in developing new scientific approaches for the design, development and evaluation of inter-vehicle communications, networking and positioning systems as part of Cooperative Intelligent Transportation Systems ensuring the safety of both roads and rail networks. This research focused on the elicitation, specification, analysis and validation of requirements for Vehicle-to-Vehicle communications and networking, and Vehicle-to-Vehicle positioning, which are accomplished with the research platform developed for this study. A number of mathematical models for communications, networking and positioning were developed from which simulations and field experiments were conducted to evaluate the overall performance of the platform. The outcomes of this research significantly contribute to improving the performance of the communications and positioning components of Cooperative Intelligent Transportation Systems.

eHealth-as-a-Service (eHaaS) : towards universal stakeholder engagement

With the introduction of the Personally Controlled Health Record (PCEHR), the Australian public is being asked to accept greater responsibility for their healthcare by taking an active role in the management of personal health information. Although well designed, constructed and intentioned, policy and privacy concerns have resulted in an eHealth model that may impact future health sharing requirements. Hence, as a case study for a consumer eHealth initative in the Australian context, eHealth-as-a-Service (eHaaS) serves as a disruptive step in in the aggregation and transformation of health information for use as real-world knowledge. The strategic value of extending the community Health Record Bank (HRB) model lies in the ability to automatically draw on a multitude of relevant data repositories and sources to create a single source of the truth and to engage market forces to create financial sustainability. The opportunity to transform the beleaguered Australian PCEHR into a realisable and sustainable technology consumption model for patient safety is explored. Moreover, the current clerical focus of healthcare practitioners acting in the role of de facto record keepers is renegotiated to establish a shared knowledge creation landscape of action for safer patient interventions. To achieve this potential however requires a platform that will facilitate efficient and trusted unification of all health information available in real-time across the continuum of care. eHaaS provides a sustainable environment and encouragement to realise this potential.

MiniOrb : a sensor interaction platform for indoor climate preferences [Extended Abstract]

We introduce the MiniOrb platform, a combined sensor and interaction platform built to understand and encourage the gathering of data around personal indoor climate preferences in office environments. The platform consists of a sensor device, gathering localised environmental data and an attached tangible interaction and ambient display device. This device allows users to understand their local environment and record preferences with regards to their preferred level of office comfort. In addition to the tangible device we built a web-based mobile application that allowed users to record comfort preferences through a different interface. This paper describes the design goals and technical setup of the MiniOrb platform.

Developing ‘design minds’ for the 21st century through a public sector initiated online design education platform

Education in the 21st century demands a model for understanding a new culture of learning in the face of rapid change, open access data and geographical diversity. Teachers no longer need to provide the latest information because students themselves are taking an active role in peer collectives to help create it. This paper examines, through an Australian case study entitled ‘Design Minds’, the development of an online design education platform as a key initiative to enact a government priority for statewide cultural change through design-based curriculum. Utilising digital technology to create a supportive community, ‘Design Minds’ recognises that interdisciplinary learning fostered through engagement will empower future citizens to think, innovate, and discover. This paper details the participatory design process undertaken with multiple stakeholders to create the platform. It also outlines a proposed research agenda for future measurement of its value in creating a new learning culture, supporting regional and remote communities, and revitalising frontline services. It is anticipated this research will inform ongoing development of the online platform, and future design education and research programs in K-12 schools in Australia.

Hybrid graphite film–carbon nanotube platform for enzyme immobilization and protection

A novel platform consisting of a multilayered substrate, activated graphite-like carbon film, and dense forest of long, vertically-aligned multiwall carbon nanotubes grown by the chemical vapor deposition is designed, fabricated, and tested for covalent immobilization of enzymatic biocatalysts with the aim of protecting them from shear forces and microbial attacks present in bioreactors. The covalent bonding ensures enzyme retention in a flow, while the dense nanotube forest may serve as a protection of the enzymes from microbial attack without impeding the flow of reactants and products. This platform was demonstrated for the two reference enzymes, horseradish peroxidase and catalase, which were immobilized without degrading their biological activity. This combination of an activated carbon layer for an efficient immobilization of biocatalysts with a protective layer of inert carbon nanotubes could dramatically improve the efficiency and longevity of enzymatic bio-catalysis employed in a large variety of advanced biotechnological processes.

Plasma-enabled graded nanotube biosensing arrays on a Si nanodevice platform : catalyst-free integration and in situ detection of nucleation events

Low-temperature plasmas in direct contact with arbitrary, written linear features on a Si wafer enable catalyst-free integration of carbon nanotubes into a Si-based nanodevice platform and in situ resolution of individual nucleation events. The graded nanotube arrays show reliable, reproducible, and competitive performance in electron field emission and biosensing nanodevices.

Self-organized Au nanoarrays on vertical graphenes : an advanced three-dimensional sensing platform

A three-dimensional surface enhanced Raman scattering (SERS)/plasmonic sensing platform based on plasma-enabled, catalyst-free, few-layer vertical graphenes decorated with self-organized Au nanoparticle arrays is demonstrated. This platform is viable for multiple species detection and overcomes several limitations of two-dimensional sensors.

A high frequency, low voltage stackable full bridge module for modular multilevel cascade converter (MMCC) research

A new small full bridge module for MMCC research is presented. Each full bridge converter cell is a single small (65 × 30 mm) multilayer PCB with two low voltage high current (22 V, 40 A) integrated half bridge ICs and the necessary isolated control signals and auxiliary power supply (2500 V isolation). All devices are surface mount, minimising cell height (4 mm) and parasitic inductance. Each converter cell can be physically stacked with PCB connectors propagating the control signals and inter-cell power connections. Many cells can be trivially stacked to create a large multilevel converter leg with isolated auxiliary power and control signals. Any of the MMCC family members is then easily formed. With a change in placement of stacking connector, a parallel connection of bridges is also possible. Operation of a nine level parallel full bridge is demonstrated at 12 V and 384 kHz switching frequency delivering a 30 W 2 kHz sinewave into a resistive load. A number of new applications for this novel module aside from MMCC development are listed.

Development of gas sensing technology for ground and airborne applications powered by solar energy : methodology and experimental results

Monitoring gases for environmental, industrial and agricultural fields is a demanding task that requires long periods of observation, large quantity of sensors, data management, high temporal and spatial resolution, long term stability, recalibration procedures, computational resources, and energy availability. Wireless Sensor Networks (WSNs) and Unmanned Aerial Vehicles (UAVs) are currently representing the best alternative to monitor large, remote, and difficult access areas, as these technologies have the possibility of carrying specialised gas sensing systems, and offer the possibility of geo-located and time stamp samples. However, these technologies are not fully functional for scientific and commercial applications as their development and availability is limited by a number of factors: the cost of sensors required to cover large areas, their stability over long periods, their power consumption, and the weight of the system to be used on small UAVs. Energy availability is a serious challenge when WSN are deployed in remote areas with difficult access to the grid, while small UAVs are limited by the energy in their reservoir tank or batteries. Another important challenge is the management of data produced by the sensor nodes, requiring large amount of resources to be stored, analysed and displayed after long periods of operation. In response to these challenges, this research proposes the following solutions aiming to improve the availability and development of these technologies for gas sensing monitoring: first, the integration of WSNs and UAVs for environmental gas sensing in order to monitor large volumes at ground and aerial levels with a minimum of sensor nodes for an effective 3D monitoring; second, the use of solar energy as a main power source to allow continuous monitoring; and lastly, the creation of a data management platform to store, analyse and share the information with operators and external users. The principal outcomes of this research are the creation of a gas sensing system suitable for monitoring any kind of gas, which has been installed and tested on CH4 and CO2 in a sensor network (WSN) and on a UAV. The use of the same gas sensing system in a WSN and a UAV reduces significantly the complexity and cost of the application as it allows: a) the standardisation of the signal acquisition and data processing, thereby reducing the required computational resources; b) the standardisation of calibration and operational procedures, reducing systematic errors and complexity; c) the reduction of the weight and energy consumption, leading to an improved power management and weight balance in the case of UAVs; d) the simplification of the sensor node architecture, which is easily replicated in all the nodes. I evaluated two different sensor modules by laboratory, bench, and field tests: a non-dispersive infrared module (NDIR) and a metal-oxide resistive nano-sensor module (MOX nano-sensor). The tests revealed advantages and disadvantages of the two modules when used for static nodes at the ground level and mobile nodes on-board a UAV. Commercial NDIR modules for CO2 have been successfully tested and evaluated in the WSN and on board of the UAV. Their advantage is the precision and stability, but their application is limited to a few gases. The advantages of the MOX nano-sensors are the small size, low weight, low power consumption and their sensitivity to a broad range of gases. However, selectivity is still a concern that needs to be addressed with further studies. An electronic board to interface sensors in a large range of resistivity was successfully designed, created and adapted to operate on ground nodes and on-board UAV. The WSN and UAV created were powered with solar energy in order to facilitate outdoor deployment, data collection and continuous monitoring over large and remote volumes. The gas sensing, solar power, transmission and data management systems of the WSN and UAV were fully evaluated by laboratory, bench and field testing. The methodology created to design, developed, integrate and test these systems was extensively described and experimentally validated. The sampling and transmission capabilities of the WSN and UAV were successfully tested in an emulated mission involving the detection and measurement of CO2 concentrations in a field coming from a contaminant source; the data collected during the mission was transmitted in real time to a central node for data analysis and 3D mapping of the target gas. The major outcome of this research is the accomplishment of the first flight mission, never reported before in the literature, of a solar powered UAV equipped with a CO2 sensing system in conjunction with a network of ground sensor nodes for an effective 3D monitoring of the target gas. A data management platform was created using an external internet server, which manages, stores, and shares the data collected in two web pages, showing statistics and static graph images for internal and external users as requested. The system was bench tested with real data produced by the sensor nodes and the architecture of the platform was widely described and illustrated in order to provide guidance and support on how to replicate the system. In conclusion, the overall results of the project provide guidance on how to create a gas sensing system integrating WSNs and UAVs, how to power the system with solar energy and manage the data produced by the sensor nodes. This system can be used in a wide range of outdoor applications, especially in agriculture, bushfires, mining studies, zoology, and botanical studies opening the way to an ubiquitous low cost environmental monitoring, which may help to decrease our carbon footprint and to improve the health of the planet.

A runtime integrity monitoring framework for real-time relative positioning systems based on GPS and DSRC

This paper provides a three-layered framework to monitor the positioning performance requirements of Real-time Relative Positioning (RRP) systems of the Cooperative Intelligent Transport Systems (C-ITS) that support Cooperative Collision Warning (CCW) applications. These applications exploit state data of surrounding vehicles obtained solely from the Global Positioning System (GPS) and Dedicated Short-Range Communications (DSRC) units without using other sensors. To this end, the paper argues the need for the GPS/DSRC-based RRP systems to have an autonomous monitoring mechanism, since the operation of CCW applications is meant to augment safety on roads. The advantages of autonomous integrity monitoring are essential and integral to any safety-of-life system. The autonomous integrity monitoring framework proposed necessitates the RRP systems to detect/predict the unavailability of their sub-systems and of the integrity monitoring module itself, and, if available, to account for effects of data link delays and breakages of DSRC links, as well as of faulty measurement sources of GPS and/or integrated augmentation positioning systems, before the information used for safety warnings/alarms becomes unavailable, unreliable, inaccurate or misleading. Hence, a monitoring framework using a tight integration and correlation approach is proposed for instantaneous reliability assessment of the RRP systems. Ultimately, using the proposed framework, the RRP systems will provide timely alerts to users when the RRP solutions cannot be trusted or used for the intended operation.

Transmedia Storytelling: From Interviewing to Multi-Platform

There is nothing new under the sun – so the saying goes, and in a digital age of recording oral histories, this holds true. Despite advances and innovations across the board in information and communication technology in the field of oral history it is essentially only the devices we record on that have changed. However, what has emerged is a plethora of ways that oral history interviews can be used to produce multimedia, or transmedia storytelling outputs- for exhibitions in public institutions, schools and by communities to engage interested groups, and in families and by individuals wanting to play with new ways of telling their family stories and histories. In 2010, QUT’s Creative Industries introduced a postgraduate unit called Transmedia Storytelling: From Interviewing to Multi-Platform, which was the first postgraduate course of its kind in Australia. Based in a Creative Writing discipline, but open to all coursework Masters, PhD, Research Masters and Doctorate of Creative Industries students, this unit introduces students to the theory and practice of semi-structured interviewing techniques, oral history conventions and applications, and the art of storytelling across various platforms.