The objective of this study was to determine the effects of feeding high concentrations of corn distillers dried grains with solubles (DDGS) on chemical composition and selected nutrient content of egg yolk. Four isocaloric diets were formulated to contain 0, 17, 35, or 50% corn DDGS. A total of two hundred forty 54-wk-old Single-Comb White Leghorn laying hens were randomly allotted to 2 birds per cage with 3 consecutive cages representing an experimental unit (EU). Each EU was randomly assigned to 1 of 4 dietary treatments according to a completely randomized design. Hens were fed for a 24-wk experimental period after transition feeding to gradually increase corn DDGS inclusion over a 4-wk period. Two sets of experimental diets were formulated to meet or exceed the NRC nutrient recommendations for laying hens. Each diet formula was fed for 12 wk. Chemical composition and nutritional components in egg yolk were measured every 2 wk. The results showed that egg yolk from hens fed a DDGS-containing diet tended to have higher fat content and lower protein content. Total polyunsaturated fatty acids were significantly increased by the DDGS diet. The contents of choline and cholesterol were initially higher in the 50% DDGS treatment group, but were not different in the later period, especially during the last 4 wk. Lutein content increased linearly as DDGS level increased. The results indicated that feeding a high level of DDGS can increase the content of lutein and polyunsaturated fatty acids in egg yolk, but may not affect the content of cholesterol and choline.