cDNA Cloning and Yeast Expression of Acyl-CoA Synthetase of Chlamydomonas reinhardtii

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.016

Abstract

Abstract: This work aims to predict and clone cDNA of Chlamydomonas reinhardtii acyl-CoA synthetase(gene cracs), and analyze its function in yeast Saccharomyces cerevisiae YB525. The cracs sequence was cloned by RT-PCR, its conserved sequence of encoded protein and phylogenetic tree were analyzed with ClustalW and MEGA6.0, then the substrate specificity in YB525 of expressed gene was analyzed. As the results, a cracs was cloned for the first time with sequence of 2 004 bp and encoded a 72.3 kD protein crACS of 667 amino acids containing two conserved regions of including acyl-CoA synthetase：the AMP-binding domain and the FACS motif. The phylogenetic tree analysis indicated that crACS shared high homology with LACs of Arabidopsis thaliana. Yeast expression experiments showed that crACS restored acyl-CoA synthetase deficient phenotype of YB525 and assimilated foreign palmitoleic acid and myristic acid. Conclusively, cracs of C. reinhardtii can activate exogenous fatty acid and belongs to acyl-CoA synthetase family.

Key words: Chlamydomonas reinhardtii, acyl-CoA synthetase, yeast YB525, phylogenetic tree, fatty acid

Song Yanzi, Jia Bin, Lin Baicheng, Hu Zhangli, Huang Ying. cDNA Cloning and Yeast Expression of Acyl-CoA Synthetase of Chlamydomonas reinhardtii[J]. Biotechnology Bulletin, 2015, 31(9): 119-124.

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