An improved HAdV-41 E1B55K-expressing 293 cell line for packaging fastidious adenovirus
► Attempt to establish a cell line to cultivate fastidious adenovirus (HAdV-41). ► Tripartite leader sequence was cloned to promote expression of HAdV-41 E1B55K gene. ► 293TE7 produced progeny HAdV-41-GFP viruses 3–15 times more than 293E12 did. ► 293TE7 was an effective packaging cell line for reco...
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Published in | Journal of virological methods Vol. 175; no. 2; pp. 188 - 196 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Kidlington
Elsevier B.V
01.08.2011
Elsevier |
Subjects | |
Online Access | Get full text |
ISSN | 0166-0934 1879-0984 1879-0984 |
DOI | 10.1016/j.jviromet.2011.05.010 |
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Summary: | ► Attempt to establish a cell line to cultivate fastidious adenovirus (HAdV-41). ► Tripartite leader sequence was cloned to promote expression of HAdV-41 E1B55K gene. ► 293TE7 produced progeny HAdV-41-GFP viruses 3–15 times more than 293E12 did. ► 293TE7 was an effective packaging cell line for recombinant or wild-type HAdV-41.
Human adenovirus type 41 (HAdV-41) is difficult to cultivate under laboratory conditions. Tripartite leader sequence (TPL) of HAdV-41 was cloned, inserted into eukaryotic expression plasmid, and used to establish a HAdV-41 E1B55K-transduced cell line (293TE7). HAdV-41 E1B55K was expressed more abundantly in 293TE7 than in 293E12, an HAdV-41 E1B55K-expressing cell line developed previously. After being infected with E1-deleted HAdV-41 vector (HAdV-41-GFP), 293TE7 synthesized more viral genomic DNA and structural proteins, which led ultimately to a significant increase of the yield of progeny viruses. Typically, 293TE7 produced progeny viruses 3–15 times more than 293E12 did, depending on the amount of seed viruses and culture time. These data demonstrated that 293TE7 was an effective packaging cell line, and implied its application for wild-type HAdV-41 isolation, HAdV-41 virological study and recombinant HAdV-41 construction. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1 |
ISSN: | 0166-0934 1879-0984 1879-0984 |
DOI: | 10.1016/j.jviromet.2011.05.010 |