Sunday, May 3, 2009
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Construction of a novel integration vector for expression of delta 6 desaturase gene in Lipomyces kononenkoae
Mulan Jiang, Xia Wan, Ping Wang, and Yinbo Zhang. Oil Crops Research Institute, CAAS, P.R.China, Xudong 2nd Road, Wuhan, China
Gamma-linolenic acid (GLA) is used as a dietary supplement and for treatment of various medical conditions. GLA is synthesized by a delta 6 fatty acid desaturase using linoleic acid (C18:2 D9,12) as a substrate. However, as the limit of its resources, to produce high levels of GLA by genetic engineering technologies is more attractive. Oil yeast Lipomyces kononenkoae, with high linoleic acid content, is an ideal expression host of delta 6 fatty acid desaturase gene compared to other expression host such as Pichia pastoris and Saccharomyces cerevisiae. To construct a novel expression vector which can be used in this oil yeast, four main components including rDNA from L. kononenkoae, hybrid promoter from pINA1296, hygromycin resistant gene and green fluorescent protein have been re-organized and inserted in the same vector. Results showed that the novel integrated expression vector has been successfully constructed. Furthermore, the delta 6 fatty acid desaturase gene from Rhizopus stolonifer has been heterogenerously expressed in L. kononenkoae. This is the first report that a novel expression vector has been constructed and proven effective in L. kononenkoae.