Establishment and analysis of transgenic clonal donor cells.

<p>(<b>A</b>) Schematic representation of n−3 fatty acid desaturase gene with linearized expression vectors. (<b>B</b>) Detection of the <i>mfat-1</i> gene in Geneticin-resistant cell clones by PCR and RT-qPCR. <i>mfat-1</i> expression vector was used as the template for positive control (PC) and untransfected syngenic cells was used as the negative control (NC). (<b>C</b>) Quantitative PCR analysis of <i>mfat-1</i> expression in positive cell clones. cDNA representing <i>mfat-1</i> was amplified with sequence specific primers. The beta-actin was used as internal control and the expression level observed in the transgenic donor cell was normalized to the value of A-3-1. (<b>D</b>) Partial gas chromatograph traces showing the polyunsaturated fatty acid profiles of total cellular lipids from the H-6-6 cells and the control cells. Note the level of n-6 polyunsaturated acids are lower whereas n−3 fatty acids are abundant in the <i>mfat-1</i> cells (right) as compared with the control cells (left), in which there is very little n−3 fatty acid.