期刊
BIOPROCESS AND BIOSYSTEMS ENGINEERING
卷 36, 期 9, 页码 1191-1197出版社
SPRINGER
DOI: 10.1007/s00449-012-0846-6
关键词
Aurantiochytrium; Ribosomal protein; Cycloheximide resistance; Gene transformation
资金
- Advanced Biomass R&D Center of Korea [2010-0029737]
- Ministry of Education, Science and Technology
- Ministry of Food, Agriculture, Forestry, and Fisheries, of the Republic Korea
In the present study, we established a genetic system for manipulating the oleaginous heterotrophic microalgae Aurantiochytrium sp. KRS101, using cycloheximide resistance as the selectable marker. The gene encoding ribosomal protein L44 (RPL44) of Aurantiochytrium sp. KRS101 was first identified and characterized. Proline 56 was replaced with glutamine, affording cycloheximide resistance to strains encoding the mutant protein. This resistance served as a novel selection marker. The gene encoding the Delta 12-fatty acid desaturase of Mortierella alpina, used as a reporter, was successfully introduced into chromosomal DNA of Aurantiochytrium sp. KRS101 via 18S rDNA-targeted homologous recombination. Enzymatic conversion of oleic acid (C18:1) to linoleic acid (C18:2) was detected in transformants but not in the wild-type strain.
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