We report the expression profile of acyl-lipid △12-desaturase （desA） gene from Synechocystis sp. PCC6803 and its effect on cell membrane lipid composition and cold tolerance in prokaryotic （Escherichia coil） and ...We report the expression profile of acyl-lipid △12-desaturase （desA） gene from Synechocystis sp. PCC6803 and its effect on cell membrane lipid composition and cold tolerance in prokaryotic （Escherichia coil） and eukaryotic （Solanum tuberosum） cells. For this purpose, a hybrid of desA and reporter gene encoding thermostable lichenase （licBM3） was constructed and used to transform these cells. The expression of this hybrid gene was measured using qualitative （Petri dish test, electrophoregram and zymogram） and quantitative methods （spectrometry and gas liquid chromatography assays）. The maximum level of linoleic acid in the bacterial cells containing hybrid gene was 1.9% of total fatty acids. Cold stress tolerance assays using plant damage index and growth parameters showed that cold tolerance was enhanced in primary transgenic lines because of increased unsaturated fatty acid concentration in their lipids. The greatest content of 18：2 and 18：3 fatty acids in primary transgenic plants was observed for lines 2 （73%） and 3 （41%）. Finally, our results showed that desaturase could enhance tolerance to cold stress in potato, and desaturase and lichenase retain their functionality in the structure of the hybrid protein where the enzymatic activity of target gene product was higher than in the case of reporter lichenase gene absence in the construction.展开更多
基金supported by the program "Biological Diversity–Gene Foundation and Genetic Diversity" of the Presidium of the Russian Academy of Sciences and the Russian Foundation for Basic Research (project nos. 08-04-90410-Ykp-a, 05-04-49186-a, 04-04-81039-Bel_a)
文摘We report the expression profile of acyl-lipid △12-desaturase （desA） gene from Synechocystis sp. PCC6803 and its effect on cell membrane lipid composition and cold tolerance in prokaryotic （Escherichia coil） and eukaryotic （Solanum tuberosum） cells. For this purpose, a hybrid of desA and reporter gene encoding thermostable lichenase （licBM3） was constructed and used to transform these cells. The expression of this hybrid gene was measured using qualitative （Petri dish test, electrophoregram and zymogram） and quantitative methods （spectrometry and gas liquid chromatography assays）. The maximum level of linoleic acid in the bacterial cells containing hybrid gene was 1.9% of total fatty acids. Cold stress tolerance assays using plant damage index and growth parameters showed that cold tolerance was enhanced in primary transgenic lines because of increased unsaturated fatty acid concentration in their lipids. The greatest content of 18：2 and 18：3 fatty acids in primary transgenic plants was observed for lines 2 （73%） and 3 （41%）. Finally, our results showed that desaturase could enhance tolerance to cold stress in potato, and desaturase and lichenase retain their functionality in the structure of the hybrid protein where the enzymatic activity of target gene product was higher than in the case of reporter lichenase gene absence in the construction.
