Successful modification of the bacterial artificial chromosome (BAC) after two-step BAC engineering is confirmed in two separate polymerase chain reactions (PCRs). The first reaction (5' co-integrate PCR) uses a forward 5' co-integrate primer (a sequence located upstream of the 5' end of the A-box) and a reverse 3' primer on the vector (175PA+50AT) or within the reporter sequence or mutated region as appropriate. The second reaction (3' co-integrate PCR) uses a forward 5' primer on the recA gene (RecA1300S) and a reverse 3' co-integrate primer (a sequence located downstream from the 3' end of the B-box). Those colonies shown to be positive in PCR analysis are further tested for sensitivity to UV light. After the resolution, colonies that have lost the excised recombination vector including sacB and recA genes become UV light sensitive.

Bacterial artificial chromosome (BAC) clones are rendered electrocompetent and transformed with the recombinant shuttle vector, pLD53SCAB/AB-box. Cointegrates are selected by growth on chloramphenicol and ampicillin to ensure recombination of the shuttle vector into the BAC.

Plasmid DNA is prepared from the recombinant shuttle vector pLD53.SCAB/A-B created by cloning of the A and B homology arms for two-step bacterial artificial chromosome (BAC) engineering. To confirm that the A-box and B-box arms have been successfully incorporated into pLD53.SCAB, the pattern of enzyme digestion of the modified plasmid is compared with that of the unmodified pLD53.SCAB. Once the shuttle vector is shown to carry the proper sequences, it is ready for transfer into the BAC host.

This protocol describes the preparation of the shuttle vector before its introduction into bacterial artificial chromosome (BAC) host cells for BAC two-step engineering. The homology arm sequences, prepared previously, are introduced by ligation into the digested shuttle vector DNA to provide sites for recombination within the BAC clone. Crude lysates of individual bacterial transformants serve as templates in polymerase chain reaction (PCR) analysis to confirm the presence of the homology arms in the recombinant shuttle vector.

The 700-bp A homology arm (A-box) and the 700-bp B homology arm (B-box) are amplified by polymerase chain reaction (PCR) using purified bacterial artificial chromosome (BAC) DNA as template for two-step BAC engineering. The resulting A-box PCR product contains an AscI site at its 5' end (the 5' primer incorporates an AscI site, and the 3' primer does not incorporate any restriction sites). The B-box PCR product contains an XmaI site at its 3' end (the 5' primer does not incorporate any restriction sites, and the 3' primer incorporates an XmaI site). The amplification products are then digested with the appropriate restriction endonucleases to render them suitable for cloning into the shuttle vector.

In two-step bacterial artificial chromosome (BAC) engineering, a single plasmid is introduced into the BAC-carrying cell lines. The shuttle vector pLD53.SCAB (or pLD53.SCAEB) carries the recA gene and the R6K origin, which requires the protein to replicate. PIR2 cells, expressing , are typically used for the amplification of the vector and maintain about 15 copies/cell of the donor vector, which is relatively stable in this host.

Tissue engineering has a unique set of tools and technologies for developing preventive strategies, diagnostics, and treatments that can play an important role during the ongoing COVID-19 pandemic.

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Repair site iFixit has shared x-ray photographs of the new Magic Keyboard for iPad Pro, and they reveal an accessory more complicated than it might appear from the outside.

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Four Nigerian marine engineering students studying in Cebu, Philippines, receive practical training and spiritual encouragement through a Logos Hope engineer.

In honor of a recently retired professor, a generous alumnus donation has enabled the creation of the Martin W. Trethewey Early Career Professorship.

The Penn State Department of Mechanical Engineering is honoring several outstanding undergraduate students through its annual awards.

Penn State senior Abigail Cowser was recently named one of 10 American Society of Civil Engineers "2020 New Faces of Civil Engineering - College Edition."

Fifteen graduating seniors from the College of Engineering have been selected to serve as student marshals for Penn State’s spring commencement ceremony, to be held virtually on May 9.

Overall, average scores were up two points since 2014 on the National Assessment of Educational Progress in Technology and Engineering Literacy.

Lipid II is an essential precursor of the bacterial cell wall biosynthesis and thereby an important target for various antibiotics. Several lanthionine-containing peptide antibiotics target lipid II with lanthionine-stabilized lipid II-binding motifs. Here, we used the biosynthesis system of the lantibiotic nisin to synthesize a two lipid II binding motifs-containing lantibiotic, termed TL19, which contains the N-terminal lipid II binding motif of nisin and the distinct C-terminal lipid II binding motif of one peptide of the two-component haloduracin (i.e. HalA1). Further characterization demonstrated that (i) TL19 exerts 64-fold stronger antimicrobial activity against E. faecium than nisin (1-22), which has only one lipid II binding site, and (ii) both the N- and C-terminal domains are essential for the potent antimicrobial activity of TL19, as evidenced by mutagenesis of each single and double domains. These results show the feasibility of a new approach to synthesize potent lantibiotics with two different lipid II binding motifs to treat specific antibiotic-resistant pathogens.

As resistance to artemisinins (current frontline drugs in malaria treatment) emerges in south East Asia, there is an urgent need to identify the genetic determinants and understand the molecular mechanisms underpinning such resistance. Such insights could lead to prospective interventions to contain resistance and prevent the eventual spread to other malaria endemic regions. Artemisinin reduced susceptibility in South East Asia (SEA) has been primarily linked to mutations in P. falciparum Kelch-13, which is currently widely recognised as a molecular marker of artemisinin resistance. However, 2 mutations in a ubiquitin hydrolase, UBP-1, have been previously associated with artemisinin reduced susceptibility in a rodent model of malaria and some cases of UBP-1 mutation variants associating with artemisinin treatment failure have been reported in Africa and SEA. In this study, we have employed CRISPR-Cas9 genome editing and pre-emptive drug pressures to test these artemisinin susceptibility associated mutations in UBP-1 in P. berghei sensitive lines in vivo. Using these approaches, we have shown that the V2721F UBP-1 mutation results in reduced artemisinin susceptibility, while the V2752F mutation results in resistance to chloroquine and moderately impacts tolerance to artemisinins. Genetic reversal of the V2752F mutation restored chloroquine sensitivity in these mutant lines while simultaneous introduction of both mutations could not be achieved and appears to be lethal. Interestingly, these mutations carry a detrimental growth defect, which would possibly explain their lack of expansion in natural infection settings. Our work has provided independent experimental evidence on the role of UBP-1 in modulating parasite responses to artemisinin and chloroquine under in vivo conditions.

When the novel coronavirus pandemic prompted the transition to remote learning at Penn State, faculty and administrators in the Department of Mechanical Engineering were prepared for the challenge.

After months of hard work and preparation, nearly a dozen Penn State Hazleton seniors are now one step closer to graduating after presenting their Capstone Research and Design Thesis projects.

Three students received scholarships in industrial and mechanical engineering from the Material Handling Education Foundation for the 2020-21 academic year.

Michael Hillman, L. Robert and Mary L. Kimball Assistant Professor of Civil and Environmental Engineering at Penn State, received a NSF CAREER Award to develop new computational methods to simulate fracture.

Mark Alpert made a generous gift $200,000 in memory of his late wife, Claire, to establish the Mark E. and Claire L. Alpert Graduate Fellowship in the Department of Civil and Environmental Engineering at Penn State.

Agency: CHR Closing Date: 6/5/2020

Differentiating good IP from mediocre or bad IP is getting more difficult, in part because it depends upon how and where it is used and in part, because even the best IP may work better in one system than another—even in chips developed by the same vendor.  

So, how do you measure IP quality and why it is so complicated?

The answer depends on who is asking. Most of the time, the definition of IP quality depends on your vantage point.  If you are an R&D manager, IP quality means something. If you are a global supply manager, IP quality means something else. If you are an SoC start-up, your measure of quality is quite different from that of an established fabless company. If you are designing IP in-house, then your considerations are very different than being a commercial IP vendor. If you are designing an automotive SoC, then we are in a totally different category. How about as an IP vendor? How do you articulate IP quality metrics to your customers?

This varies greatly by the type of IP, as well. When it comes to interface (hard) IP and controllers, if you are an R&D manager, your goal is to design IP that meets the IP specifications and PPA (power, performance, and area) targets. You need to validate your design via silicon test chips. This applies to all hard PHYs, which must be mapped to a particular foundry process. For controllers that are in RTL form—we called these soft IP—you have to synthesize them into a particular target library in a particular foundry process in order to realize them in a physical form suitable for SoC integration. Of course, your design will need to go through a series of design validation steps via simulation, design verification and passing the necessary DRC checks, etc. In addition, you want to see the test silicon in various process corners to ensure the IP is robust and will perform well under normal process variations in the production wafers.

For someone in IP procurement, the measure of quality will be based on the maturity of the IP. This involves the number of designs that have been taped out using this IP and the history of bug reports and subsequent fixes. You will be looking for quality of the documentation and the technical deliverables. You will also benchmark the supplier’s standard operating procedures for bug reporting and technical support, as well as meeting delivery performance in prior programs. This is in addition to the technical teams doing their technical diligence.

An in-house team that is likely to design IP for a particular SoC project will be using an established design flow and will have legacy knowledge of last generation’s IP. They may be required to design the IP with some reusability in mind for future programs. However, such reusability requirements will not need to be as stringent and as broad as those of commercial IP vendors because there are likely to be established metrics and procedures in place to follow as part of the design team’s standard operating procedures. Many times, new development based on a prior design that has been proven in use will be started, given this stable starting point. All of these criteria help the team achieve a quality outcome more easily.

Then, if designing for an automotive SoC, additional heavy lifting is required.  Aside from ensuring that the IP meets the specifications of the protocol standards and passes the compliance testing, you also must pay attention to meeting functional safety requirements. This means adherence to ISO 26262 requirements and subsequently achieving ASIL certification. Oftentimes, even for IP, you must perform some AEC-Q100-related tests that are relevant to IP, such as ESD, LU, and HTOL.

To read more, please visit: https://semiengineering.com/why-ip-quality-is-so-difficult-to-determine/

Today, we are living in the era where whatever we think of as an idea is not far from being implemented…thanks to machine learning (ML) and artificial intelligence (AI) entering into the...

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•Lockheed Martin deploys the 3DEXPERIENCE platform as an engineering and manufacturing planning toolset •Multi-year collaboration aims to speed timelines and improve efficiencies of next generation products •Digital experience platform approach drives advances in complex, sophisticated aircraft innovation

Buildings account for nearly four-tenths of U.S. energy consumption through heating, cooling and other electricity use, according to the Energy Information Administration. And if that energy comes from fossil fuels, it releases more greenhouse gases that drive human-caused climate change.

Buildings account for nearly four-tenths of U.S. energy consumption through heating, cooling and other electricity use, according to the Energy Information Administration. And if that energy comes from fossil fuels, it releases more greenhouse gases that drive human-caused climate change.

Naval shipbuilder Navantia Australia, a subsidiary of Spain-based Navantia S.A., has opened a new design and engineering centre in Melbourne.

Buildings account for nearly four-tenths of U.S. energy consumption through heating, cooling and other electricity use, according to the Energy Information Administration. And if that energy comes from fossil fuels, it releases more greenhouse gases that drive human-caused climate change.

Europe’s utilities are re-evaluating their business models due to the energy transition. Members of POWER-GEN Europe’s Advisory Board consider how a reliance on fossil fuels is no longer politically desirable, forcing utilities to transform their portfolios to adapt to radical change.

In 1984, when this year’s POWER-GEN 2015 Woman of the Year Kim Greene started engineering school, about 16 percent of her class was made up of women. Today, more than 30 years later, that number has jumped to just 18 percent.

Greene, the Chief Operating Office of Southern Company was one of three finalists for the 2015 Woman of the Year award during Power Generation Week and on Tuesday December 8, she took part in a panel discussion with her co-finalists: Terry Jester, CEO and Chairman of Silicor Materials and Roxann Laird, Director of the National Carbon Capture Center, during the Women in Power luncheon.

The fact that women only make up 18 percent of engineering students is an alarming statistic and was the topic of discussion for much of the panel. To combat it, Greene believes that everyone should be encouraging young women to study engineering. “Fathers, mothers, uncles, aunts, school teachers,” she said, explaining that it should be important to all of us to encourage girls to pursue education in technical fields that will (hopefully) ultimately lead to careers in the energy industry. Laird explained that she does simple things in her household, such as conducting science experiments at birthday parties for her daughter.

Also important is persistence, said Greene. Girls may need a bit of extra encouragement when they do poorly on a test or fail a class. If a mentor can offer words of encouragement like “keep your chin up” and “you can do this,” that may be all that is needed to keep a girl motivated who may otherwise have become discouraged.

As far as rising through the ranks of an organization, Jester believes it isn’t about being the smartest person in the room or always making the right decisions. Women just need to believe that they are capable of being charge. If you make a mistake, own it and move on, said Greene. It’s ok to admit you are wrong, don’t dig in your heels.

Panelists were also asked for tips on how to re-enter the workplace after a hiatus or after switching careers and all agreed on one thing: “don’t apologize for the gap.” Jester believes that someone who has re-dedicated herself to the workplace makes for an excellent employee.

At the end of the day “engineers solve problems,” said Laird adding, “it’s a fun industry.” Jester said that there are misperceptions in the industry that engineers are stuck in a cube or an office doing very boring jobs but it is not like that. Those of us in the industry know its fun.

Robynn Andracsek, an engineer with Burns & McDonnell and member of the Women in Power committee led the panel discussion during the luncheon.

The purpose of the Women in Power committee is to provide role models to inspire young women to pursue careers in energy. Anyone in the energy industry can nominate a woman for the Woman of the year award. Nominations are open from April to August each year.

What better way to learn about the electronics industry than from real people successfully pursuing their careers in great companies? Charlene Gunter du Plessis, Senior Director of the IPC Education Foundation talked to Kate Stees, a Materials and Process Engineer at Lockheed Martin Missiles & Fire Control in Ocala, Florida. Kate currently works in the […]

Buildings account for nearly four-tenths of U.S. energy consumption through heating, cooling and other electricity use, according to the Energy Information Administration. And if that energy comes from fossil fuels, it releases more greenhouse gases that drive human-caused climate change.

Samsung Engineering said on August 16 that it won two petrochemical power generation projects worth 550 billion won jointly with PTT Public Company Ltd., a Thai state-run oil company. This has been less than a week since the Korean company announced a large-scale oil refinery building project in Oman worth more than 1 trillion won. The project calls for building a petrochemical plant and an electricity generation plant in Rayong, Thailand's largest industrial city 150 kilometers southeast of ...