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dc.contributor.authorLin, Chi-Hung
dc.contributor.authorJarvis, Donald L
dc.date.accessioned2021-05-26T04:00:12Z
dc.date.available2021-05-26T04:00:12Z
dc.date.issued2013
dc.identifier.issn0168-1656
dc.identifier.doi10.1016/j.jbiotec.2013.02.007
dc.identifier.urihttp://hdl.handle.net/10072/404695
dc.description.abstractGenetically transformed lepidopteran insect cell lines have biotechnological applications as constitutive recombinant protein production platforms and improved hosts for baculovirus-mediated recombinant protein production. Insect cell transformation is often accomplished with a DNA construct(s) encoding a foreign protein(s) under the transcriptional control of a baculovirus immediate early promoter, such as the ie1 promoter. However, the potential utility of increasingly stronger promoters from later baculovirus gene classes, such as delayed early (39K), late (p6.9), and very late (polh), has not been systematically assessed. Hence, we produced DNA constructs encoding secreted alkaline phosphatase (SEAP) under the transcriptional control of each of the four temporally distinct classes of baculovirus promoters, used them to transform insect cells, and compared the levels of SEAP RNA and protein production obtained before and after baculovirus infection. The ie1 construct was the only one that supported SEAP protein production by transformed insect cells prior to baculovirus infection, confirming that only immediate early promoters can be used to isolate transformed insect cells for constitutive recombinant protein production. However, baculovirus infection activated transgene expression by all four classes of baculovirus promoters. After infection, cells transformed with the very late (polh) and late (p6.9) promoter constructs produced the highest levels of SEAP RNA, but only low levels of SEAP protein. Conversely, cells transformed with the immediate early (ie1) and delayed early (39K) promoter constructs produced lower levels of RNA, but equal or higher levels of SEAP protein. Unexpectedly, the 39K promoter construct provided tightly regulated, baculovirus-inducible protein production at higher levels than the later promoter constructs. Thus, this study demonstrated the utility of the 39K promoter for insect cell engineering, particularly when one requires higher levels of effector protein production than obtained with ie1 and/or when constitutive transgene expression adversely impacts host cell fitness and/or genetic stability. © 2013 Elsevier B.V.
dc.description.peerreviewedYes
dc.languageEnglish
dc.publisherElsevier
dc.relation.ispartofpagefrom11
dc.relation.ispartofpageto17
dc.relation.ispartofissue1
dc.relation.ispartofjournalJournal of Biotechnology
dc.relation.ispartofvolume165
dc.subject.fieldofresearchBiological sciences
dc.subject.fieldofresearchEngineering
dc.subject.fieldofresearchcode31
dc.subject.fieldofresearchcode40
dc.subject.keywordsScience & Technology
dc.subject.keywordsLife Sciences & Biomedicine
dc.subject.keywordsBiotechnology & Applied Microbiology
dc.subject.keywordsBaculovirus-insect cell system
dc.subject.keywordsGenetic transformation
dc.titleUtility of temporally distinct baculovirus promoters for constitutive and baculovirus-inducible transgene expression in transformed insect cells
dc.typeJournal article
dc.type.descriptionC1 - Articles
dcterms.bibliographicCitationLin, C-H; Jarvis, DL, Utility of temporally distinct baculovirus promoters for constitutive and baculovirus-inducible transgene expression in transformed insect cells, Journal of Biotechnology, 2013, 165 (1), pp. 11-17
dcterms.dateAccepted2013-02-15
dcterms.licensehttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.date.updated2021-05-26T03:55:53Z
dc.description.versionAccepted Manuscript (AM)
gro.rights.copyright© 2013 Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence (http://creativecommons.org/licenses/by-nc-nd/4.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, providing that the work is properly cited.
gro.hasfulltextFull Text
gro.griffith.authorLin, Chi-Hung


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