scholarly journals Rhythmic ATP release from the cyanobacterial circadian clock protein KaiC revealed by real‐time monitoring of bioluminescence using firefly luciferase

2020 ◽  
Author(s):  
Risa Mutoh ◽  
Keita Iwata ◽  
Takahiro Iida ◽  
Masahiro Ishiura ◽  
Kiyoshi Onai
Keyword(s):  
Circadian Clock ◽  
Real Time ◽  
Atp Release ◽  
Firefly Luciferase ◽  
Real Time Monitoring ◽  
Clock Protein

Rhythmic post-transcriptional regulation of the circadian clock protein mPER2 in mammalian cells: A real-time analysis

2006 ◽  
Vol 401 (1-2) ◽  
pp. 44-48 ◽  
Author(s):  
Keigo Nishii ◽  
Iori Yamanaka ◽  
Maya Yasuda ◽  
Yota B. Kiyohara ◽  
Yoko Kitayama ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Circadian Clock ◽  
Real Time ◽  
Mammalian Cells ◽  
Time Analysis ◽  
Real Time Analysis ◽  
Post Transcriptional Regulation ◽  
Clock Protein

Firefly Luciferase Inhibitor-Conjugated Peptide Quenches Bioluminescence: A Versatile Tool for Real Time Monitoring Cellular Uptake of Biomolecules

2013 ◽  
Vol 25 (1) ◽  
pp. 4-10 ◽  
Author(s):  
Pekka K. Poutiainen ◽  
Teemu Rönkkö ◽  
Ari E. Hinkkanen ◽  
Jorma J. Palvimo ◽  
Ale Närvänen ◽  
...  
Keyword(s):  
Real Time ◽  
Cellular Uptake ◽  
Firefly Luciferase ◽  
Real Time Monitoring ◽  
Versatile Tool

scholarly journals Real-Time Monitoring of Chloroplast Gene Expression by a Luciferase Reporter: Evidence for Nuclear Regulation of Chloroplast Circadian Period

2006 ◽  
Vol 26 (3) ◽  
pp. 863-870 ◽  
Author(s):  
Takuya Matsuo ◽  
Kiyoshi Onai ◽  
Kazuhisa Okamoto ◽  
Jun Minagawa ◽  
Masahiro Ishiura
Keyword(s):  
Gene Expression ◽  
Circadian Clock ◽  
Real Time ◽  
Nuclear Genome ◽  
Firefly Luciferase ◽  
Luciferase Reporter ◽  
Chloroplast Gene ◽  
Chloroplast Gene Expression ◽  
Period Gene ◽  
Circadian Expression

ABSTRACT Chloroplast-encoded genes, like nucleus-encoded genes, exhibit circadian expression. How the circadian clock exerts its control over chloroplast gene expression, however, is poorly understood. To facilitate the study of chloroplast circadian gene expression, we developed a codon-optimized firefly luciferase gene for the chloroplast of Chlamydomonas reinhardtii as a real-time bioluminescence reporter and introduced it into the chloroplast genome. The bioluminescence of the reporter strain correlated well with the circadian expression pattern of the introduced gene and satisfied all three criteria for circadian rhythms. Moreover, the period of the rhythm was lengthened in per mutants, which are phototactic rhythm mutants carrying a long-period gene in their nuclear genome. These results demonstrate that chloroplast gene expression rhythm is a bona fide circadian rhythm and that the nucleus-encoded circadian oscillator determines the period length of the chloroplast rhythm. Our reporter strains can serve as a powerful tool not only for analysis of the circadian regulation mechanisms of chloroplast gene expression but also for a genetic approach to the molecular oscillator of the algal circadian clock.

scholarly journals Real-time monitoring of extracellular ATP in bacterial cultures using thermostable luciferase

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0244200
Author(s):  
Julian Ihssen ◽  
Nina Jovanovic ◽  
Teja Sirec ◽  
Urs Spitz
Keyword(s):  
Real Time ◽  
Susceptibility Testing ◽  
Bacterial Species ◽  
Atp Release ◽  
Extracellular Atp ◽  
Clinical Diagnostics ◽  
Broth Culture ◽  
Current Evidence ◽  
Real Time Monitoring ◽  
Bacterial Cultures

Adenosine triphosphate (ATP) is one of the most important indicators of cell viability. Extracellular ATP (eATP) is commonly detected in cultures of both eukaryotic and prokaryotic cells but is not the focus of current scientific research. Although ATP release has traditionally been considered to mainly occur as a consequence of cell destruction, current evidence indicates that ATP leakage also occurs during the growth phase of diverse bacterial species and may play an important role in bacterial physiology. ATP can be conveniently measured with high sensitivity in luciferase-based bioluminescence assays. However, wild-type luciferases suffer from low stability, which limit their use. Here we demonstrate that an engineered, thermostable luciferase is suitable for real-time monitoring of ATP release by bacteria, both in broth culture and on agar surfaces. Different bacterial species show distinct patterns of eATP accumulation and decline. Real-time monitoring of eATP allows for the estimation of viable cell number by relating luminescence onset time to initial cell concentration. Furthermore, the method is able to rapidly detect the effect of antibiotics on bacterial cultures as Ampicillin sensitive strains challenged with beta lactam antibiotics showed strongly increased accumulation of eATP even in the absence of growth, as determined by optical density. Patterns of eATP determined by real-time luminescence measurement could be used to infer the minimal inhibitory concentration of Ampicillin. Compared to conventional antibiotic susceptibility testing, the method presented here is faster and more sensitive, which is essential for better treatment outcomes and reducing the risk of inducing antibiotic resistance. Real-time eATP bioluminescence assays are suitable for different cell types, either prokaryotic or eukaryotic, thus, permitting their application in diverse fields of research. It can be used for example in the study of the role of eATP in physiology and pathophysiology, for monitoring microbial contamination or for antimicrobial susceptibility testing in clinical diagnostics.

scholarly journals 2PT230 ATP release from cyanobacterial circadian clock protein KaiC(The 50th Annual Meeting of the Biophysical Society of Japan)

2012 ◽  
Vol 52 (supplement) ◽  
pp. S143-S144
Author(s):  
Keita Iwata ◽  
Risa Mutoh ◽  
Kiyoshi Onai ◽  
Masahiro Ishiura
Keyword(s):  
Circadian Clock ◽  
Annual Meeting ◽  
Atp Release ◽  
Biophysical Society ◽  
Clock Protein

scholarly journals Real-time monitoring of circadian clock oscillations in primary cultures of mammalian cells using Tol2 transposon-mediated gene transfer strategy

2010 ◽  
Vol 10 (1) ◽  
pp. 3 ◽  
Author(s):  
Kazuhiro Yagita ◽  
Iori Yamanaka ◽  
Noriaki Emoto ◽  
Koichi Kawakami ◽  
Shoichi Shimada
Keyword(s):  
Gene Transfer ◽  
Circadian Clock ◽  
Real Time ◽  
Mammalian Cells ◽  
Primary Cultures ◽  
Real Time Monitoring ◽  
Tol2 Transposon

1618: Real-Time Monitoring of Nitric Oxide and Blood Flow During Ischemia-Reperfusion in the Rat Testis

2006 ◽  
Vol 175 (4S) ◽  
pp. 521-521
Author(s):  
Motoaki Saito ◽  
Tomoharu Kono ◽  
Yukako Kinoshita ◽  
Itaru Satoh ◽  
Keisuke Satoh
Keyword(s):  
Nitric Oxide ◽  
Blood Flow ◽  
Real Time ◽  
Ischemia Reperfusion ◽  
Real Time Monitoring ◽  
Rat Testis

GaN heteroepitaxy by remote plasma MOCVD : Real time monitoring by laser reflectance interferometry

2001 ◽  
Vol 11 (PR3) ◽  
pp. Pr3-1175-Pr3-1182 ◽  
Author(s):  
M. Losurdo ◽  
A. Grimaldi ◽  
M. Giangregorio ◽  
P. Capezzuto ◽  
G. Bruno
Keyword(s):  
Real Time ◽  
Real Time Monitoring ◽  
Remote Plasma ◽  
Laser Reflectance
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