2,666 research outputs found

    Monomeric PcrA helicase processively unwinds plasmid lengths of DNA in the presence of the initiator protein RepD

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    The helicase PcrA unwinds DNA during asymmetric replication of plasmids, acting with an initiator protein, in our case RepD. Detailed kinetics of PcrA activity were measured using bulk solution and a single-molecule imaging technique to investigate the oligomeric state of the active helicase complex, its processivity and the mechanism of unwinding. By tethering either DNA or PcrA to a microscope coverslip surface, unwinding of both linear and natural circular plasmid DNA by PcrA/RepD was followed in real-time using total internal reflection fluorescence microscopy. Visualization was achieved using a fluorescent single-stranded DNA-binding protein. The single-molecule data show that PcrA, in combination with RepD, can unwind plasmid lengths of DNA in a single run, and that PcrA is active as a monomer. Although the average rate of unwinding was similar in single-molecule and bulk solution assays, the single-molecule experiments revealed a wide distribution of unwinding speeds by different molecules. The average rate of unwinding was several-fold slower than the PcrA translocation rate on single-stranded DNA, suggesting that DNA unwinding may proceed via a partially passive mechanism. However, the fastest dsDNA unwinding rates measured in the single-molecule unwinding assays approached the PcrA translocation speed measured on ssDNA

    Playing Reindeer Games: Native Alaskans and the Federal Trust Doctrine

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    The Reindeer Industry Act of 1937 established a de facto Native Alaskan monopoly in the reindeer industry as a means of subsistence that would allow Native Alaskans to remain self-sufficient and continue to practice their traditional customs. In 1997, the Ninth Circuit held that the Reindeer Act did not preclude nonNatives from owning and selling reindeer, thereby opening the reindeer industry up to non-Natives. The unique Native Alaskan culture of the Seward Peninsula, which depends upon the reindeer industry, is in jeopardy as a result of competition it now faces from non-Natives. The federal government has a fiduciary obligation to protect the cultural welfare of Native Alaskans as a result of the trust relationship that exists between the federal government and Native Americans. The federal government has yet to take action to fulfill that fiduciary obligation to the Native Alaskan reindeer herders of the Seward Peninsula

    Technical writing versus technical writing

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    Two terms, two job categories, 'technical writer' and 'technical author' are discussed in terms of industrial and business requirements and standards. A distinction between 'technical writing' and technical 'writing' is made. The term 'technical editor' is also considered. Problems inherent in the design of programs to prepare and train students for these jobs are discussed. A closer alliance between industry and academia is suggested as a means of preparing students with competent technical communication skills (especially writing and editing skills) and good technical skills

    As One We Will Never Be

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