Omega-3 Polyunsaturated Fatty Acid Dehydrogenase Gene Fatl In Optimizing The Expression Of Fibroblast Cells Of Mice

Long chainω-3 polyunsaturated fatty acids (PUFAs) such as EPA and DHA are important for many normal physiological functions of mammals. In contrast, over dose ofω-6 PUFAs intake had been reported to have many adverse effects for mammal’s health.ω-3 PUFAs have been demonstrated can counteract the abnormal lipid metabolism of excessiveω-6 PUFAs intake. Long chain co-3 PUFAs are derived from the substrate alpha linolenic acid (a-LNA 18:3 co-3) and the substrates are metabolized to longer chain derivatives in some kinds of algae and nematode. Mammals lack the corresponding co-3 desaturase responsible for synthesizing long chainω-3 PUFAs in vivo. So the intake ofω-3 PUFAs is recommended for its health benefits to protect against heart disease, diabetes, and potentially cancer. But as a result of the increased consumption of foods rich inω-6 fatty acids (milk, butter, animal fat, vegetable oil) and the reduced consumption of foodstuff rich in co-3 fatty acids (sea fish or nuts), there has caused a imbalance of co-6 PUFAs:ω-3 PUFAs ratios highly exceeded 5:1, the rational dietary ratio that health studies suggested. In this research, we cloned the cDNA of co-3 fatty acid desaturase enzyme (fatl gene) from Caenorhabditis elegans. As the fatl gene under study is a kind of heterologous protein for mammal, in order to acquire a high-level production of the desired protein we optimized some rare codons under 10% utilization rate in accordance with codon usage table for Mus musculus. We hope the alteration would make it more suitable for mammalia gene expression. Then the recombinant vectors were transfected into mouse embryonic fibrocyte cell. Results showed cells transfected by pEGFPC1-fatl and pEGFPCl-Ofatl both can convert theω-6 PUFAs toω-3 PUFAs effectively. The cellular fatty acids composition analysis demonstrated that the optimized gene seemed exert a better function of co-3 PUFAs desaturase in vitro. Cell apoptosis analysis showed that the wild type and optimized fatl gene both can counteract the growth inhibition adversity of overdoseω-6 PUFAs and compared to the control group, they showed a lower cell apoptosis rate. Our conclusion hopely could lay a foundation for future transgenic research about this highly concernedω-3 PUFAs desaturase gene.