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| Cloning and Expression crtI Gene in Methylobacterium sp MB200 |
| LIN Qiong-shan1,2,3, TANG Xian-lai1,4, LI Xian1,2,3, JIANG Cheng-jian1,2,3, LI Jun-fang1,2,3, SHEN Pei-hong 1,2,3 |
1. College of Life Science and Technology of Guangxi University, Nanning 530005, China;
2. The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, Nanning 530005, China;
3. State Key Laboratory for Conservation and Utilization of Agricultural Bioresources in the Subtropics, Nanning 530005, China;
4. The Science and Techenology Department of Guangxi, Nanning 530005, China |
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Abstract The partial mutants library of M. sp MB200 was constructed by transposon mutagenesis and 11552 mutants were obtained which could survived in medium with methanol as the sole carbon source. Rescreening results showed that 33 strains were colorless mutants. The crtI gene was got from a colorless mutant by molecular cloning techniques. It shared 93% identity with the crtI gene of M. populi BJ001, M. chloromethanicum CM4 and M. extorquens AM1 at the nucleotide level reported in Genbank. The gene size is 1539 bp, encoding the phytoene desaturase (CrtI) that catalyzes phytoene to form lycopene with four desaturations, or catalyzes phytoene to form neurosporene with three desaturations, therefore it plays an important role in the biosynthetic pathway of carotenoid. Here, a recombinant plasmid pCM80-crtI was constructed by ligating the crtI to the vector pCM80, then imported into M. sp MB200 to obtain recombinant strain MB200/pCM80-crtI. The activity of enzyme from MB200/pCM80-crtI was more by 40% than M. sp MB200. The experimental results provided a theoretical reference to improve the biosynthetic and metabolic pathways of carotenoid in Methylobacterium.
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Received: 20 March 2012
Published: 25 August 2012
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