Gene Cloning of the Orange Carotenoid Protein from Cyanobacteria,and Its Ectopic Expression and Functional Evaluation in Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2016.07.021

Abstract

Abstract: The aim is to verify the orange carotenoid protein（OCP）in cyanobacteria can scavenge singlet oxygen（¹O₂）independent of carotenoids in vivo. The cyanobacterial OCP gene was amplified by PCR,and then subjected to prokaryotic inducible expression and further evaluation of anti-¹O₂ activity in Escherichia coli by bacterial dot-plating test and growth curve assay. Results showed that the gene（SyOCP）of OCP was successfully amplified from the metagenome of the cyanobacteria water,with a coding region of 954 bp. Bioinformatics analysis indicated that SyOCP was from the major cyanobacteria species—Synechocystis sp. PCC 6803. By IPTG induction,the SyOCP gene was highly expressed in E. coli with soluble protein. In addition,the ectopic expression of SyOCP in recombinant E. coli resulted in the notable enhancement of it on the tolerance to the common ¹O₂-induced photosensitizer dye,methylene blue. The intrinsic anti-¹O₂role of cyanobacterial OCP（without binding carotenoids）was in vivo verified in E. coli.

Key words: cyanobacteria, orange carotenoid protein, metagenome, singlet oxygen, methyl blue

LIU Chang LUO Zhu ZHANG Meng-ru YANG Yu-mei LIU Xian GONG Ming ZOU Zhu-rong. Gene Cloning of the Orange Carotenoid Protein from Cyanobacteria,and Its Ectopic Expression and Functional Evaluation in Escherichia coli[J]. Biotechnology Bulletin, 2016, 32(7): 138-145.

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