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Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.bbrc.2004.02.148
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dc.titleHigh-level expression of a codon optimized recombinant dust mite allergen, Blo t 5, in Chinese hamster ovary cells
dc.contributor.authorLim, L.H.
dc.contributor.authorLi, H.Y.
dc.contributor.authorLee, B.W.
dc.contributor.authorChua, K.Y.
dc.contributor.authorCheong, N.
dc.date.accessioned2013-11-18T08:31:43Z
dc.date.available2013-11-18T08:31:43Z
dc.date.issued2004
dc.identifier.citationLim, L.H., Li, H.Y., Lee, B.W., Chua, K.Y., Cheong, N. (2004). High-level expression of a codon optimized recombinant dust mite allergen, Blo t 5, in Chinese hamster ovary cells. Biochemical and Biophysical Research Communications 316 (4) : 991-996. ScholarBank@NUS Repository. https://doi.org/10.1016/j.bbrc.2004.02.148
dc.identifier.issn0006291X
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/47990
dc.description.abstractBlo t 5 is a major allergen from house dust mite Blomia tropicalis. Purification of native Blo t 5 (nBlo t 5) from whole dust mite extract is tedious and gave low yield. In this study, we demonstrated that codon optimization facilitated high-level expression of Blo t 5 in Chinese hamster ovary (CHO)-K1 cells and thus allows production of sufficient recombinant cBlo t 5 for specific immunotherapy. A codon optimized Blo t 5 gene was synthesized by PCR and the codon optimized or wild-type Blo t 5 gene in pcDNA3.0 was transfected into CHO-K1 cells and stably selected with Geneticin (G418). Western-immunoblot analysis of spent culture media detected a positive band at 14kDa for the codon optimized but not wild-type gene transfectants. In addition, a stable CHO-K1 clone produced up to 13mg/L of the cBlo t 5 protein having a high correlation of human IgE reactivities and allergenicity to the native Blo t 5, thus indicating proper conformation of this recombinant form. © 2004 Elsevier Inc. All rights reserved.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1016/j.bbrc.2004.02.148
dc.sourceScopus
dc.subjectBlomia tropicalis, Blo t 5
dc.subjectChinese hamster ovary cells, CHO
dc.subjectCodon optimization
dc.subjectHouse dust mite
dc.typeArticle
dc.contributor.departmentPAEDIATRICS
dc.description.doi10.1016/j.bbrc.2004.02.148
dc.description.sourcetitleBiochemical and Biophysical Research Communications
dc.description.volume316
dc.description.issue4
dc.description.page991-996
dc.description.codenBBRCA
dc.identifier.isiut000220579300004
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