Abstract:
:The entire cloned human adenovirus type 5 (Ad5) genome is known to be able to generate infectious virus after transfection into 293 cells when the both ends of the genome are exposed by digestion with appropriate restriction enzymes. However, when one or both ends of the genome are tagged with nucleotides and are not intact, whether the tagged end of the viral genome was remained tagged or corrected to be intact during the generation of viral clones has been unclear and, if such oligonucleotide removal occurs, how does the virus remove these tagged sequences and thereby restore its proper structure? Here, we show in our semi-quantitative study that the generation efficiency of virus clones decreases depending on the length of nucleotide tags at the both ends and that both the oligonucleotide tags were precisely removed during virus generation with restoration of the proper terminal sequences. Interestingly the viral genome of which one end was tagged, while the other was attached about 12-kb sequences, did generate intact viral clones at a reduced but significant efficiency. From these results, we here propose a possible mechanism whereby the terminal-protein-deoxycytidine complex enters from the enzyme-cleaved end and reaches deoxyguanine at the initiating position of DNA synthesis in vivo. A replication origin at one end, embedded deeply in double-stranded DNA, can be activated by two cycles of one-directional full-length DNA synthesis initiated by the other exposed replication origin about 30 kilobases away. We also describe new cassette cosmids which can use not only Pac I but also Bst BI for construction of an adenovirus vector, without reducing construction efficiency.
journal_name
Microbiol Immunoljournal_title
Microbiology and immunologyauthors
Fukuda H,Terashima M,Koshikawa M,Kanegae Y,Saito Idoi
10.1111/j.1348-0421.2006.tb03829.xsubject
Has Abstractpub_date
2006-01-01 00:00:00pages
643-54issue
8eissn
0385-5600issn
1348-0421pii
JST.JSTAGE/mandi/50.643journal_volume
50pub_type
杂志文章abstract::Congenitally athymic BALB/cA nu/nu mice were employed to elucidate the role of the thymus in experimental Mycoplasma pulmonis strain m53 infection, and nu/+ mice were used for comparison. Chronic polyarthritis was frequently produced in both of nu/nu and nu/+ mice by intravenous injection of the organisms. Macroscopic...
journal_title:Microbiology and immunology
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doi:10.1111/j.1348-0421.1979.tb00537.x
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journal_title:Microbiology and immunology
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journal_title:Microbiology and immunology
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journal_title:Microbiology and immunology
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pub_type: 杂志文章
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journal_title:Microbiology and immunology
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journal_title:Microbiology and immunology
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