期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 437, 期 1, 页码 23-28出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2013.06.026
关键词
Coelenterazine; Furimazine; Codon-optimized gene; Gaussia; Reporter assay
资金
- Kurata Grants
- Naito Foundation
- JSPS KAKENHI Grant [23710266, 24651257]
- Grants-in-Aid for Scientific Research [23710266, 24651257] Funding Source: KAKEN
The codon-optimized gene for the mutated 19 kDa protein (nanoKAZ), which is the catalytic component of Oplophorus luciferase, was expressed in Escherichia coli cells and the recombinant protein was highly purified. The secretory expression of nanoKAZ from CHO-K1 cells was performed by fusing the secretory signal peptide sequence of Gaussia luciferase to the amino-terminus of nanoKAZ. The substrate specificity for the purified nanoKAZ and the nanoKAZ secreted into the cultured medium was determined, indicating that his-coelenterazine (bis-CTZ) and newly synthesized 6h-f-coelenterazine (6h-f-CTZ) are an efficient substrate for the glow luminescence reaction of nanoKAZ. (c) 2013 Elsevier Inc. All rights reserved.
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