Establishment and comparison of three different codon optimization of fat1 gene in transgenic mammary epithelial cell lines
Fat1 gene is an n-3 fatty acid encoding desaturase from Caenorhabditis elegan (C. elegan). It can raise the n-3/n-6 polyunsaturated fatty acids (PUFAs) ratio in mammalian cells. To reveal the impact of different codon optimizations of fat1 gene in influencing the catalysis efficiency of n-6 PUFAs into n-3 PUFAs in mammalian cells, the fat1 gene from C. elegan coding sequence was optimized based on the codon usage frequency preference of bovine muscle protein, milk protein and the mixtype, named as rfat1, nfat1 and yfat1, respectively. Three different codon optimized fat1 gene mammalian cell expression vectors were constructed and transfected into mammary epithelial cells of Chinese Holstein Cows. As expected, the specific fragment was amplified by RT-PCR from the transfected cells. Also, different catalysis efficiencies were observed with different codon optimizations. This study will help to understand the impact of codon usage bias, and the mechanisms of tissue specific regulation of gene expression of fat1 on bovine mammary epithelial cell lines.
Key words: Fat1, codon usage bias, gene expression, mammary epithelial cells.