K. intermedius ENS15 gDNA was sequenced at Novogene Europe (Cambridge, UK) using the Illumina Novaseq 6000 system. The raw reads were quality controlled using FastQC18, trimmed using Trimmomatic19 and de-novo assembled using SPAdes20. The misassembles were identified using QUAST (5.0.2)21. Contig reordering, missassembly correction and gap filling was conducted with GFinisher22 tool using K. intermedius AF2 genome (GenBank accession: GCA_000817255.2) as the reference. The scaffolds were manually linked, the genome completeness was analysed by CheckM tool23, and annotated using Prokka24. Phylogenetic relatedness and genome-based taxonomy was performed in Type Strain Genome Server (TYGS)25 ( , accessed on 23.07.2021). The plasmid sequences in the raw reads and draft genome were identified using plasmidSPAdes26 and Recycler27, respectively. The genome assembly can be found in the NCBI GenBank under the accession number GCA_000964425.1.
Digital Control System Benjamin C Kuo Pdf Free 51
AHL-, ATc-, IPTG- and arabinose-inducible systems, under the control of pLux, pTet, pLac and pBAD promoters, have been previously characterized in Komagataeibacter spp5,6. In the studies, the pLux promoters demonstrated highest mRFP1 expression and less leakiness than the tested counterparts. Due to the lack of lac repressors in Komagataeibacter genomes, pLac promoter was employed as a constitutive promoter by Teh et al.6. Furthermore, in the study, constructs carrying arabinose-inducible pBAD promoter was studied in Komagataeibacter spp. In comparison to pLux promoter, the authors identified that the arabinose-inducible construct imparted low mRFP1 expression and identified that the presence of arabinose in the growth medium negatively affected nanocellulose production. Contemplating the literature reports, we investigated mRFP1 expression in K. intermedius ENS15 using previously characterized AHL (pLux)- and ATc (pTet)-inducible systems, and a Cumate-inducible expression system that has never been characterized in Komagataeibacter spp. (Fig. 5). The Cumate switch consisting of a cymR repressor gene and CuO repressor-binding operator under strong constitutive promoters have been employed for tight expression of recombinant genes in E. coli strains and Bacillus spp33,54. However, we noted that the Cumate-inducible system was quite leaky in K. intermedius ENS15 (Fig. 5a) and the inducer negatively impacted the cell growth (Table S6). Such effect on the cell growth was not observed in K. intermedius ENS15 cells harboring AHL- and ATc-inducible constructs, except in the highest ATc concentration (10 µg/ml) which could be attributed towards the tetracycline toxicity (Table S6). In the case of mRFP1 induction using AHL- and ATc-inducible systems in K. intermedius ENS15, contradictory results to that published for K. rhaeticus iGEM strain was observed. Although AHL-inducible construct imparted low leakiness, a higher induction was observed using ATc-inducible construct with an optimal inducer concentration of 0.5 µg/ml (Fig. 5b). Furthermore, robust induction of mRFP1 expression while encased in the pellicle indicates that, despite entrapped within nanocellulose, the K. intermedius ENS15 cells can effectively receive signals from their environment enabling its applicability in applications requiring long-term cell survival and tolerance to toxic compounds (Fig. 5d,e).
Blockchain technology can be used to create a permanent, public, transparent ledger system for compiling data on sales, tracking digital use and payments to content creators, such as wireless users[120] or musicians.[121] The Gartner 2019 CIO Survey reported 2% of higher education respondents had launched blockchain projects and another 18% were planning academic projects in the next 24 months.[122] In 2017, IBM partnered with ASCAP and PRS for Music to adopt blockchain technology in music distribution.[123] Imogen Heap's Mycelia service has also been proposed as a blockchain-based alternative "that gives artists more control over how their songs and associated data circulate among fans and other musicians."[124][125]
As an example, we can consider a smart home system, which consists of many sensors and actuators. The actuators are used to lock/unlock the doors, switch on/off the lights or other electrical appliances, alert users of any threats through alarms or notifications, and control the temperature of a home (via a thermostat).
A typical IoT application has many producers of data (sensors), a set of servers to process the data, and a set of actuators. FiWare refers to the information collected by sensors as context information. It defines generic REST APIs to capture the context from different scenarios. All the context information is sent to a dedicated service called a context broker. FiWare provides APIs to store the context and also query it. Moreover, any application can register itself as a context consumer, and it can request the context broker for information. It also supports the publish-subscribe paradigm. Subsequently, the context can be supplied to systems using context adapters whose main role is to transform the data (the context) based on the requirements of the destination nodes. Moreover, FiWare defines a set of SNMP APIs via which we can control the behavior of IoT devices and also configure them. 2ff7e9595c
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