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Title: High-level expression of a codon optimized recombinant dust mite allergen, Blo t 5, in Chinese hamster ovary cells
Authors: Lim, L.H. 
Li, H.Y.
Lee, B.W. 
Chua, K.Y. 
Cheong, N.
Keywords: Blomia tropicalis, Blo t 5
Chinese hamster ovary cells, CHO
Codon optimization
House dust mite
Issue Date: 2004
Citation: Lim, 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.
Abstract: Blo 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.
Source Title: Biochemical and Biophysical Research Communications
ISSN: 0006291X
DOI: 10.1016/j.bbrc.2004.02.148
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