In:
Luminescence, Wiley, Vol. 29, No. 1 ( 2014-02), p. 13-19
Abstract:
Expression of bacterial luciferase enzyme ( lux ) in eukaryotic cells would provide a new bioreporter system for in vivo imaging and diagnostics technology. In spite of this, until now only a few efforts have been made to express bacterial luciferase enzyme in eukaryotic cells. We attempted to synthesize an expression construct of luxA and luxB genes from Vibrio fischeri . The luxA and luxB genes were cloned into the MCS of pTZ57R via the 5' kpn I, Bam HI and Bam HI, Eco RI restriction sites to generate pTZ57R/luxA and pTZ57R/luxB respectively, then newly synthesized constructs were cleaved with the same enzymes and respectively cloned into the pcDNA3.1 + (Hyg) and pcDNA3.1 + (Neo) expression vectors to create pcDNA3.1 + (Hyg)/luxA and pcDNA3.1 + (neo)/luxB. Recombinant constructs were cotransfected to the NIH3T3 cell line. Gene expression was confirmed by reverse transcription–polymerase chain reaction, sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blotting; in addition, bioluminescence characteristics of transfected NIH3T3 cell lines were evaluated by decanal supplement. In conclusion, in the current research, separate vector systems were constructed, which are composed of bacterial luciferase genes ( luxA and luxB ) that accordingly have not already been reported. These results hold promise toward the potential development of an autonomous light‐generating lux reporter system in eukaryotic cells. Copyright © 2013 John Wiley & Sons, Ltd.
Type of Medium:
Online Resource
ISSN:
1522-7235
,
1522-7243
Language:
English
Publisher:
Wiley
Publication Date:
2014
detail.hit.zdb_id:
2001819-8
SSG:
12
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