Graduate Thesis Or Dissertation

 

Adenovirus complementary strand synthesis in vivo Public Deposited

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/1n79h775d

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  • Replication of plasmid molecules containing a single adenovirus terminal sequence and specially arranged inverted repeat sequences were studied. When the plasmids were linearized to expose the adenovirus terminal sequence and transfected into 293 cells with helper adenovirus DNA, a larger molecule was detected. The new molecule contained adenovirus terminal sequences at both ends of the molecule. This approach provides a sensitive assay to determine which adenovirus terminal sequences are required in cis for replication Plasmids carrying deletion mutations within cloned adenovirus terminal sequences have been studied to map the replication origin. The replication origin is contained entirely within the first 67 by of the adenovirus inverted terminal repeat (ITR). This region could be further subdivided into two functional domains: a minimal replication origin and an adjacent auxilliary region which boosted the efficiency of replication by more than 100- fold. The minimal origin occupies the first 18 to 21 by and includes sequences conserved between all adenovirus serotypes. The adjacent auxilliary region extends past nucleotide 36 but not past nucleotide 67, and contains the binding sites for nuclear factor I and nuclear factor III. Complementary strand replication has been studied in vivo by using plasmid molecules containing a single adenovirus replication origin and different sizes of inverted repeat sequences. Inverted repetitious sequences 33 by or longer are required for complementary strand replication. Complementary strand synthesis cannot be detected when plasmids contain inverted repeat sequences 28 by or smaller.
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