In:
Intervirology, S. Karger AG, Vol. 55, No. 5 ( 2012), p. 349-355
Abstract:
〈 b 〉 〈 i 〉 Objective: 〈 /i 〉 〈 /b 〉 In order to gain further insight into the function of the enteric adenovirus short fiber (SF), we have constructed a recombinant dodecahedron containing the SF protein of HAdV-41 and the HAdV-3 〈 i 〉 penton 〈 /i 〉 base. 〈 b 〉 〈 i 〉 Methods: 〈 /i 〉 〈 /b 〉 Recombinant baculoviruses expressing the HAdV-41 SF protein and HAdV-3 〈 i 〉 penton 〈 /i 〉 base were cloned and amplified in 〈 i 〉 Sf 〈 /i 〉 9 insect cells. Recombinant dodecahedra were expressed by coinfection of High Five™ cells with both baculoviruses, 72 h post-infection. Cell lysate was centrifuged on sucrose density gradient and the purified recombinant dodecahedra were recovered. 〈 b 〉 〈 i 〉 Results: 〈 /i 〉 〈 /b 〉 Analysis by negative staining electron microscopy demonstrated that chimeric dodecahedra made of the HAdV-3 〈 i 〉 penton 〈 /i 〉 base and decorated with the HAdV-41 SF were successfully generated. Next, recombinant dodecahedra were digested with pepsin and analyzed by Western blot. A ‘site-specific’ proteolysis of the HAdV-41 SF was observed, while the HAdV-3 〈 i 〉 penton 〈 /i 〉 base core was completely digested. 〈 b 〉 〈 i 〉 Conclusion: 〈 /i 〉 〈 /b 〉 These results show that, in vitro, the HAdV-41 SF likely undergoes proteolysis in the gastrointestinal tract, its natural environment, which may facilitate the recognition of receptors in intestinal cells. The results obtained in the present study may be the basis for the development of gene therapy vectors towards the intestinal epithelium, as well as orally administered vaccine vectors, but also for the HAdV-41 SF partner identification.
Type of Medium:
Online Resource
ISSN:
0300-5526
,
1423-0100
Language:
English
Publisher:
S. Karger AG
Publication Date:
2012
detail.hit.zdb_id:
1482863-7
SSG:
12
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