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.

Using a cross-country sample of bank-dependent public fi rms we study the international spillovers of a change in banking regulation on corporate borrowing. For identi cation we examine how US rms' liabilities vis-a-vis banks, non-bank lenders and bond markets evolve after an increase in capital requirements implemented by the European Banking Authority (EBA) in 2011. We find that US firms experience a reduction in credit lines but not in term loans from EU banks.

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.

Recent literature has highlighted that international trade is mostly priced in a few key vehicle currencies, and is increasingly dominated by intermediate goods and global value chains (GVCs). Taking these features into account, this paper reexamines the business cycle dynamics of international trade and its relationship with monetary policy and exchange rates.

Our ongoing celebration of the Library of Congress National Book Festival continues with crime and mystery writer Karin Slaughter discussing "Broken," part of her Will Trent series, on the Fiction & Mystery stage at the 2010 Festival.

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

No global real estate market despite higher price synchronisation and growing role of international investors, central banks find (Press release, 18 February 2020)

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

The last time Charlene Chiddenton saw her father in person was on March 7 — visitor restrictions at long-term care homes during the pandemic had kept them apart — and less than two months later, she was saying goodbye to her father by video call.

Holland America and Princess cruise lines issued notices this week cancelling the remainder of the season in Atlantic Canada, citing global health concerns, and that's taking a huge bite out of the Port of Sydney's budget.

A Dalhousie University scientist and a NSCAD textile professor are teaming up to create a new fabric. It could be used to locally manufacture personal protective equipment like surgical masks. As the CBC's Colleen Jones reports, they have received a COVID-19 grant to study the idea.

A number of fishing zones in the Gulf of St. Lawrence have been closed after North Atlantic right whales were sighted earlier this week.

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.

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

A 9-year-old girl was found safe in the Lanaudière region, after she had gone missing Thursday evening in the woods behind her family's second home. 

Asylum seekers make up a large portion of the "guardian angels" Quebec Premier François Legault has praised in his daily COVID-19 briefings: the orderlies who have no guarantee they'll be allowed to stay in Canada.

While the STM is strongly urging passengers to wear a face mask or face covering at all times, it won't be mandatory, said chair Philippe Schnobb.

Hope you didn't pack away those winter boots yet: southwestern Manitoba, from Riding Mountain south to the Canada-U.S. border, is expected to get heavy rain and snow Friday night.

Winnipeg police officers who arrested a man who then went into medical distress and later died were not at fault, an investigation by Manitoba's police watchdog has found.

Eight COVID-19 cases are now connected to that workplace, including six employees.

The union that represents thousands of Manitoba workers is asking what safety protocols were in place when a construction worker was killed after a trench wall collapsed on him earlier this week.

We can expect to see more seasonable temperatures by next weekend, says a meteorologist from Environment Canada.

With the business world -- and the world at large, for that matter -- changing at what feels like a moment's notice, businesses and brands have never been required to be as limber as in this current moment. Marketing leaders want hard evidence and objective facts for decision making. It wasn't long ago that multi-touch attribution was the prized child of the hype cycle among marketers.

Oracle and Zoom just entered a deal that for once is more about technological audacity than about dollars -- a partnership to host Zoom on Oracle Cloud Infrastructure. In just a few months -- basically since the beginning of the novel coronavirus pandemic -- Zoom has seen demand for its service grow from about 10 million daily meeting participants to more than 300 million.

Consumers and companies worldwide have ramped up online ordering for software products and digital goods as they struggle to improve productivity and security while working remotely and spending more time at home. The sharp spike in online commerce aligns with the timing of the current global pandemic. Software-based offerings accounted for the highest levels of growth.

Health PEI plans to create a COVID-19 unit within the Prince Edward Home to be used for any long-term care resident — living in any long-term care facility on the Island — who is diagnosed with the virus.

RCMP in Souris, P.E.I., say an incident that shut down the community from 6 p.m. Friday until close to midnight ended with one man in custody and no injuries.

Artists from Michigan and around the world are painting 50 murals in Flint to refocus the city's image on art rather than the lead-tainted water crisis.

Authorities investigating Flint's water crisis have used search warrants to seize from storage the state-owned mobile devices of former Gov. Rick Snyder and 65 other current or former officials, The Associated Press has learned.

Prosecutors stunningly dropped all criminal charges Thursday against eight people in the Flint water scandal and pledged to start the investigation from scratch.

All remaining criminal charges of city and state officials stemming from the Flint Water Crisis have been dismissed.

Prosecutors who dropped charges against eight people in the Flint water scandal explained their decision in a public forum Friday night, telling frustrated, shocked and saddened residents they must look at hundreds of mobile devices and millions of documents that a previous investigative team never reviewed.

A former FBI agent who investigated the Flint water scandal says the team was moving toward new charges when new prosecutors took over.

Former Michigan Gov. Rick Snyder says he won’t continue with a fellowship at Harvard University following backlash over his role in Flint’s water crisis.

The Flint Registry is holding in-person enrollment across the city of Flint all week. Officials are trying to track the health of residents made ill by the Flint Water Crisis.

Nearly 5 ½ years ago, the water source for the city of Flint was switched in an effort to save money. A disaster followed resulting in deaths, illness and mistrust of government. Tuesday at 10:00 p.m. on WKAR-TV, the documentary series Frontlin e examines the crisis by bringing to light conversations and documents never seen on television before. We spoke with Abby Ellis, the Michigan native who is the film’s director.

The Flint Water Crisis captured national attention about the dangers of lead contamination in public water supplies. Now, nearly six years after the problem was first detected, health officials are reporting on the well-being of children in Flint.

Suddenly, the White House loves Bernie Sanders.

Global cross-border bank claims continued to expand rapidly, growing at 9% year on year. As in previous quarters, the expansion was mainly due to claims on the non-bank sector, which grew at 12% year on year. The growth in claims on non-bank financial institutions was particularly strong (+17%). European banks' cross-border lending, which went through a prolonged contraction after the Great Financial Crisis (GFC) of 2007-09, has been expanding again since the start of 2018.

The annual growth rate of global cross-border bank claims fell to 6%, down from 9% at end-September 2019. The short-term share of foreign banks' claims, a key indicator of external vulnerability, is elevated for a number of borrowing emerging market and developing economies (EMDEs). Borrowers in EMDEs had untapped credit lines of $610 billion, or roughly 10% of the stock of global foreign claims on EMDEs at end-2019. Saudi Arabia joined the reporting population for the locational banking statistics (LBS) with data starting from Q4 2017, bringing the number of countries reporting these data to 48.

Interview with Ms Isabel Schnabel, Member of the Executive Board of the European Central Bank, in To Vima (Greek daily newspaper), conducted by Mr Angelos Athanasopoulos and published on 4 April 2020.

Interview with Mr Luis de Guindos, Vice-President of the European Central Bank, and La Vanguardia, conducted by Mr Manel Pérez and published on 12 April 2020.

Interview with Ms Christine Lagarde, President of the European Central Bank, and Le Parisien, conducted by Mr Matthieu Pelloli and published on 9 April 2020.

Interview with Ms Christine Lagarde, President of the European Central Bank, and France Inter, conducted by Mr Ali Baddou and Ms Carine Bécard on 9 April 2020.

Remarks by Benoît Cœuré, Head of the Bank for International Settlements Innovation Hub, at the Reinventing Bretton Woods Committee - Chamber of Digital Commerce webinar on "The world economy transformed", 17 April 2020.