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Higher intensity luciferase and pH sensor GFP


Higher intensity luciferase

Brighter than commonly used luciferase!

These luciferases have at most four-time maximum luminous intensity than that of widely used luciferase of the Photinus pyralis, a common North American firefly. These luciferases as a reporter, allow to monitor gene expression with high sensitivity and time-lapse observation.

Observation of luminescence of transfected HeLa cells.
HeLa cells were transfected with expression vectors harbouring luciferase coding regions, respectively. Luminescence was observed under the condition of 2 mM D-luciferin. Left, PmatLuc1, DkumLuc1, MALuci2Luc. Right, SflaRE1Luc, PsagRE1Luc.

 

Catalog no. Name of clone Origin Intracellular luminescence intensity(vie, Phothinus pyralis) Maximal absorption wavelength (pH8, in vitro) misc.
25℃ 37℃
RDB14359 pPmatLuc1 Pyrocoeli matsumurai 4 times or more 560 nm 567 nm luciferase cDNA is cloned in pUC19 vector.
RDB14360 pDkumLuc1 Drilaster kumejimensis 4 times 558 nm 562 nm
RDB14363 pMALuci2Luc Malaysian Luciola sp. (1) 2 times 556 nm 557 nm
RDB14362 pSflaRE1Luc Stenocladius flavipennis 2 times 608 nm 609 nm
RDB14361 pPsagRE1Luc Pyrocoeli sagulatus 2 times 605 nm 606 nm

1The luciferase gene originated from Malaysian firefly was developed together with Perak State Development Corporation and Nimura Genetic Solutions in conformity with the Convention on Biological Diversity.

Multiple colors with one substrate!

Green-, Yellow- or Red-emission can be obtained by using D-luciferin as a substrate of each luciferase. Simultaneous monitoring of two genes can be allowed by using green- and red-luciferases.

High stabilities under various pH conditions!

These luciferases are less susceptible under variety of pH or temperature and the stable results can be obtained. Please visit our web site to see results of luminescence at 25℃ and 37℃.

Purified recombinant luciferases were mixed with GTA Buffer (final conc. 200 mM), ATP (final conc. 2 mM), MgSO4 (final conc. 2 mM), D-Luciferin (final conc. 1 mM) and luminescence under the different pH conditions were observed. Left, 25℃. Right, 37℃.
PmatLuc1
DkumLuc1
MALuci2Luc
PsagRE1Luc
SflaRE1Luc

Akiyoshi R, Ogoh K, Suzuki H. 2012. Firefly luciferase. Japan patent unregistered 2012-235756
Akiyoshi R, Ogoh K, Suzuki H. 2013. Firefly luciferase. Japan patent unregistered 2013-74861
Ogoh K, Akiyoshi R, Suzuki H. 2013. Firefly luciferase. Japan patent unregistered 2013-138668
Ogoh K, Akiyoshi R, Suzuki H. 2013. Firefly luciferase. Japan patent unregistered 2013-81459
Akiyoshi R, Ogoh K, Suzuki H. 2015. FIREFLY LUCIFERASE, Patent publication US2015/0291938 A1

 

Thermostable GFP


Upper, before heating. Lower, after heating at 100℃ for 10 minutes.
Observed at room temperature.
Relative fluorescent intencity.
  25℃ 70℃ 100℃
GFP of A. victoria 1072 1087 629
WT 4770 4229 703
Th1wt 8067 7236 2162
Th2wt 7668 7525 3711

 

Catalog no. Name of clone Characteristic misc.
RDB14364 pCoGFP-Th1wt wild type GFP cDNA is cloned
in pUC19 vector.
RDB14366 pCoGFP-Th2wt wild type
RDB14365 pCoGFP-Th1co Optimize codons for transfection into mammalian cells
RDB14367 pCoGFP-Th2co Optimize codons for transfection into mammalian cells

Murai M, Ogoh K, Kinebuchi T, Suzuki H. Thermostable fluorescent protein and its utility. Japan patent unregistered 2014-60947

 

pH sensor GFP

Ogoh, K. et al. Dual-color-emitting green fluorescent protein from the sea cactus Cavernularia obesa and its use as a pH indicator for fluorescence microscopy. Luminescence 28 (4): 582-591, 2013.

 

Catalog no. Name of clone Characteristic misc.
RDB14368 pCoGFP-wt wild type GFP cDNA is cloned
in pUC19 vector.
RDB14369 pCoGFP-mam From blue to green at pH 5 – 6
RDB14370 pCoGFP-V0 From blue to green at pH 5 – 6
RDB14371 pCoGFP-V1 From blue to green at pH 6 – 7
RDB14372 pCoGFP-V2 From blue to green at pH 7 – 8
RDB14373 pCoGFP-V3 From blue to green at pH 9 – 10
RDB14374 pCoGFP-V4 From blue to green at pH 9 – 10

(N.N. 2016.11.16)

2017.01.06