CC-5163 H5-M1 mt+ (high lipid producing)

$30.00

From Rasha Abdrabu, NYU Abu Dhabi, October 2015

UV mutagenesis of CC-503 yielding high lipid producing isolate.

CC-503 was grown using TAP liquid media. 3.25 x 10^8 cells/mL were transferred to TAP agar plates exposed to ultraviolet (UV) light at a distance of 30 cm (253.7 nm, 100 μW/cm2, 60 Hz, NuAire, http://www.nuaire.com) for 2 minutes under sterile conditions. The plates were subsequently kept in dark for one day to prevent photo reactivation of the DNA repair mechanism. The plates were then allowed to grow under light for approximately a week. The resulting colonies were suspended in TAP liquid media and stained by BODIPY 505/515 for visualizing cells containing neutral lipids, which were then sorted using BD FACSAria III instrument. Using this protocol in an iterative fashion, an increase in lipid accumulation after four rounds of mutagenesis was observed. The cells selected after the fourth round, were sorted again without mutagenesis to further enrich the lipid accumulating cells. The sorted cells from the forth round of mutagenesis were grown using a TAP liquid media. The resulting single colonies were picked and suspended in TAP liquid media and stained by BODIPY 505/515 to illuminate cells containing neutral lipids, which were then analyzed using a FACS instrument. H5-M1 produced the highest labeled signal.

Rasha Abdrabu, Sudhir Kumar Sharma, Basel Khraiwesh, Kenan Jijakli, David R. Nelson, Amnah Alzahmi, Joseph Koussa, Mehar Sultana, Sachin Khapli, Ramesh Jagannathan, Kourosh Salehi-Ashtiani. "Single-Cell Characterization of Microalgal Lipid Contents with Confocal Raman Microscopy". Essentials of Single-Cell Analysis; part of the series Series in BioEngineering pp 363-382: 22 January 2016