A dual fluorescence protein expression system detects cell cycle dependent protein noise

被引:0
|
作者
Murugesan, Akshaya [1 ,2 ,5 ]
Alshagrawi, Reshod A. [3 ]
Thiyagarajan, Ramesh [4 ]
Kandhavelu, Meenakshisundaram [1 ,2 ]
机构
[1] Tampere Univ, Fac Med & Hlth Technol, Mol Signaling Grp, POB 553, Tampere 33101, Finland
[2] BioMediTech, POB 553, Tampere 33101, Finland
[3] King Saud Univ, Coll Food Sci & Agr, Dept Food Sci & Nutr, Riyadh, Saudi Arabia
[4] Prince Sattam Bin Abdulaziz Univ, Coll Med, Dept Basic Med Sci, Al Kharj 11942, Saudi Arabia
[5] Madurai Kamaraj Univ, Lady Doak Coll, Dept Biotechnol, Madurai 625002, India
关键词
Protein expression; Noise components; Cell cycle; Promoter dynamics; ESCHERICHIA-COLI; RNA; STOCHASTICITY; MAGNESIUM; DYNAMICS; IONS; MG2+;
D O I
10.1016/j.ijbiomac.2024.130262
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Inherently identical cells exhibit significant phenotypic variation. It can be essential for many biological processes and is known to arise from stochastic, 'noisy', gene expression that is determined by intrinsic and extrinsic components. It is now obvious that the noise varies as a function of inducer concentration. However, its fluctuation over the cell cycle is limited. Applying dual colour fluorescence protein reporter system, Cyan Fluorescent Protein (CFP) and Yellow fluorescent protein (YFP) tagged multi-copy plasmids, we determine variation of the noise components over the phases in lac promoter induced by Isopropyl beta-D-1-thiogalactopyranoside (IPTG) and in presence of additional Magnesium, Mg2+ ion. We, also, estimate the how such system deviates from observations of single-copy plasmid. Found 25 % difference between multi-copy system and single-copy system clarifies that observed noise is considerable and estimates population behaviour during the cell cycle. We show that total variation in cells induced with IPTG is determined by higher extrinsic than intrinsic noise. It increases from Lag to Exponential phase and decreases from Retardation to Stationary phase. By observing slow and fast dividing cells, we show that 5 mM Mg2+ increases population homogeneity compared to 2.5 mM Mg2+ in the environment. The experimental data obtained using dual colour fluorescence protein reporter system demonstrates that protein expression noise, depending on intra cellular ionic concentration, is tightly controlled by phase of the cell.
引用
收藏
页数:7
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