Chlamydomonas sp. ICE-L, an Antarctic ice alga, has high tolerance ability to freezing and salinity. Glutathione (GSH) is an important small antioxidative molecule in the growth and stress responses of plants including algae. | Turkish Journal of Botany Turk J Bot (2018) 42: 371-381 © TÜBİTAK doi: Research Article Characteristic of Glutamate Cysteine Ligase Gene and its response to the salinity and temperature stress in Chlamydomonas sp. ICE-L from Antarctica 1 1, 1 1 1 2 Yue-yue PENG , Yu DING * , Xiao-chen TANG , Ji-chang JIAN , Jin-hui WANG , Quan-fu WANG 1 Guangdong Provincial Key Lab of Pathogenic Biology and Epidemiology for Aquatic Economic Animals; Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education Institutions; Fisheries College, Guangdong Ocean University, Zhanjiang, Guangdong, P. R. China 2 Ocean College, Harbin Institute of Technology, Weihai, . China Received: Accepted/Published Online: Final Version: Abstract: Chlamydomonas sp. ICE-L, an Antarctic ice alga, has high tolerance ability to freezing and salinity. Glutathione (GSH) is an important small antioxidative molecule in the growth and stress responses of plants including algae. We cloned a full-length cDNA encoding glutamate cysteine ligase (ICE-LGCL), the key enzyme of GSH synthesis, from Chlamydomonas sp. ICE-L by RT-PCR and rapid amplification of cDNA ends technique (RACE). The cDNA has 2199 bp nucleotides with an open reading frame (ORF) of 1452 bp encoding a polypeptide of 453 amino acids. BLASTP algorithm results showed that ICE-LGCL shared 51%–70% amino acid sequence identity with the reported GCLs and shared the highest identity with Chlamydomonas reinhardtii. We also successfully made ICE-LGCL protein express in E. coli BL21. The optimum expression conditions are the induced reagent IPTG of mmol/L, temperature of 37 °C, and the induced time of 4 h. The expression patterns of ICE-LGCL in mRNA by real-time PCR analysis showed that ICE-LGCL expressed under the different temperature and salinity challenges. Low temperature and low salinities stimulated .
