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.

CPMI report: Payment aspects of financial inclusion in the fintech era, April 2020

FSI Briefs No 4, April 2020. Currently, insurers are more likely to experience losses from financial market volatility than from higher insurance claims arising from Covid-19. Few insurance supervisors have seen a need to strengthen or adjust prudential requirements to insulate insurers from current financial market uncertainties. So far, authorities have responded mainly by taking measures to provide operational relief to insurers from regulatory and supervisory requirements so that they can continue providing insurance services. These measures will also help insurers to enhance risk monitoring of their Covid-19 financial exposures. Some authorities have set out expectations for insurers to conserve capital through prudent exercise of dividend and variable remuneration policies. The aim is to enhance their resilience against huge uncertainties from potential Covid-19 fallout. Other capital-related measures should relieve supervisory pressures and reduce the tendency of insurers to manage their investments in a procyclical manner. These measures include: extending the supervisory intervention ladder, triggering the countercyclical lever and recalibrating capital requirements. The far-reaching impact of Covid-19 calls for sustained vigilance by both supervisors and insurers. In the post-pandemic phase, the extraordinary measures currently warranted will need to be unwound through a carefully crafted exit strategy that preserves sound risk management practices and protects policyholders' interests.

This paper reviews post-crisis financial regulatory reforms, examines how they fit together and identifies open issues. Specifically, it takes stock of the salient new features of bank and CCP international standards within a unified analytical framework.

Rural educator Frank Cyr had the vision and pull to force the nation to standardize the color of the ubiquitous vehicle

Open for visitors, these houses model upcycling at its finest

British product designer Lucy Hughes has invented a biodegradable plastic made from fish offcuts

From gas stations to public libraries, these celebrity architect-designed buildings are worth a road trip

Press release about the Basel Committee publishing consultation paper on revisions to the credit valuation adjustment risk framework, 28 November 2019.

BIS Press Release - Easing trade tensions lift sentiment: BIS Quarterly Review, 8 December 2019

Press release: Basel Committee invites comments on the design of a prudential treatment for crypto-assets, 12 December 2019.

Press release about the Basel Committee launching the consolidated Basel Framework, 16 December 2019.

The BIS Board of Directors has decided to invite the central banks of Kuwait, Morocco and Vietnam to become members of the BIS. This is the first such expansion since 2011 and will take the number of members to 63.

Basel Committee Press release "Basel Committee meets to review vulnerabilities and emerging risks, advance supervisory initiatives and promote Basel III implementationl", 27 February 2020.

BIS Press Release - Shaping the future of payments: BIS Quarterly Review, 1 March 2020

BIS statement, 16 March 2020.

Press release about Basel Committee assesses members' implementation of the Net Stable Funding Ratio and large exposures framework, 19 March 2020

Press release "Governors and Heads of Supervision announce deferral of Basel III implementation to increase operational capacity of banks and supervisors to respond to Covid-19", 27 March 2020

BCBS Press release "Basel Committee sets out additional measures to alleviate the impact of Covid-19", 3 April 2020

BCBS Press release "Basel Committee and IOSCO announce deferral of final implementation phases of the margin requirements for non-centrally cleared derivatives", 3 April 2020

CPMI Press release "Jurisdictions move towards full implementation of standards for financial market infrastructures", 8 April 2020.

Press release "Saudi G20 Presidency and the Bank for International Settlements (BIS) Innovation Hub invite global innovators to solve RegTech and SupTech challenges", 27 April 2020

BCBS Press release "Basel Committee issues progress report on banks' implementation of the 'Principles for effective risk data aggregation and reporting'", 29 April 2020

Rutgers' RUCDR Infinite Biologics received an amended emergency use authorization from the FDA late Thursday for the first SARS-CoV-2 coronavirus test that will allow people to collect their own saliva at home and send to a lab for results.

The American Geriatrics Society annually honors researchers, clinicians, educators, and emerging health professionals who have made outstanding contributions to high-quality, person-centered care for older people.

A protein shredder that occurs in cell membranes of brain cells apparently also indirectly regulates the fat metabolism.

When we find something particularly beautiful or impressive, we literally get big eyes: Our pupils dilate. The pupil controls how much light enters the eye and falls on the retina.

Classified released a music video for his song, Good News, this week featuring a pack of kids hanging out with seniors at a continuing care home in Enfield, N.S. It was filmed before the COVID-19 outbreak in the province.

Graduation is on hold for Dalhousie University medical student Manveer Bal, so he's decided to spend his newfound free time volunteering at the epicentre of the COVID-19 outbreak in Nova Scotia.

People in Truro, N.S., are reeling after the disappearance of three-year-old Dylan Ehler as the community tries to come to grips with another potential tragedy in the area.

A woman who lost her sister to COVID-19 at the start of the month is thanking the staff at Northwood who took "excellent" care of her in her final days. Jean Harrigan, 90, is being remembered for her caring and lively nature.

As Nova Scotians gazed up at the Canadian Forces Snowbirds soaring through the clouds last weekend, Matt MacKenzie was looking down and feeling the weight of successive tragedies that had enveloped his home province. 

Carmen Figueroa Sotelo has been interpreting all the news conferences for English audiences since the beginning of the pandemic in Quebec

Quebecers, like the rest of the world, are growing accustomed to the uncertainty that's accompanied the pandemic. But they may not appreciate their government adding to that already hefty burden.

The COVID-19 crisis has wreaked havoc on Quebec's economy, with the province recording its highest unemployment rate since at least 1976, when the data was first tracked.

For close to 15 years, Reema Singh has been baking and selling cakes from her tiny shop on Parc Avenue in Montreal's Mile End. Cocoa Locale has been open throughout the pandemic because — well, it turns out that cake is essential.

The current deconfinement plan in the Montreal area could lead to a rapid increase in deaths in the city, according to findings by Quebec's public health institute. 

The COVID-19 pandemic has created a surge in nationwide shipments of specialities from legendary restaurants

The Metuchen Farmers Market, like many others, has moved to online orders and drive-thru pickups during the coronavirus pandemic

Live professional baseball games could be televised in the United States as early next week, with South Korea's Yonhap News Agency reporting Monday that ESPN and the Korea Baseball Organization are nearing an agreement.

Major League Baseball could hold a rescheduled opening day between mid-June and July 4 and play at least 80 games this season, The Athletic's Ken Rosenthal reported Tuesday.

Derek Jeter, Larry Walker and the rest of this year's Baseball Hall of Fame class will have to wait another year for their big moment at Cooperstown.

Vowing to return next summer, this year's Little League World Series was cancelled due to the coronavirus pandemic. All seven World Series tournaments and their respective regional qualifying events are off this summer.

Cleveland Indians reliever Emmanuel Clase was suspended 80 games by Major League Baseball on Friday for testing positive for a banned performance-enhancing drug.

Baseball Hall of Famer Andre Dawson spoke to CBC's John Northcott about the realities of running a funeral home during the pandemic.