mCherry fluorescence correlation with western blot band intensity

Overview

The results present in this dataset were obtained from recombinant Chlamydomonas reinhardtii expressing the fluorescent protein mCherry in different constructs formats, aiming to recombinant protein secretion. 

Image present in this dataset were obtained by following the steps described in the protocol dx.doi.org/10.17504/protocols.io.kfpctmn. 

Fluorescence measurements were obtained by growing cc1690 in 50 mL TAP media on a rotary shaker, set to 150 rpm, under constant illumination (50 μmol photons/m²s). Then, 100 μL of each strain was transferred to a clear bottom 96-well plate (Corning Costar, Tewksbury, MA, USA) and fluorescence was measured using an Infinite® M200 PRO plate reader (Tecan, Männedorf, Switzerland). Fluorescence was measured at excitation 575/9 nm and emission 608/20 nm. Supernatant samples were obtained by spinning 1 mL sample at 15000 × g for 10 min and transferring 100 μL from each well to the clear bottom 96-well plate (Corning Costar, Tewksbury, MA, USA), followed by fluorescence measurement.

Files Info

021115_20171219 Correlation_Fluore_vs_Densito_mCherry_50mL_7_days.xlsx   -> data points for fluorescence measurements and pixel intensity obtained with FiJI (Fiji Is Just ImageJ). 

022615 WB SP mCHerry Lysate 3.JPG -> Raw western blot image for lysate samples

022615 WB SP mCHerry Supernatant 3 .JPG -> Raw western blot image for supernatant samples 

20171219 Fluor_Densito_Correlation_Lys.png -> Correlation plot of fluorescence and pixel intensity for lysate samples

WB_Lysa_7days_for_Densitom.png  -> Prepared image for densitometry analysis of lysate samples

20171219 WB_Sup_7days_for_Densitom.png -> Correlation plot of fluorescence and pixel intensity for supernatant samples

WB_Sup_7days_for_Densitom.png  -> Prepared image for densitometry analysis of supernatant samples

Consider citing our work. 

Molino JVD, de Carvalho JCM, Mayfield SP (2018) Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii. PLoS ONE 13(2): e0192433. https://doi.org/10.1371/journal. pone.0192433