In this article, flour with gradient relative content of high-molecular-weight glutenin sub-units (HMW-GS) is obtained by mixing the flour of transgenic line with completely silent HMW-GS and wild line, with their dough quality measured. The results show that the gluten formation could not be detected when the relative HMW-GS content was less than 16%. If the content of HMW-GS is 16–24%, the soft gluten could be formed, but the gluten with great force could not be detected. When HMW-GS relative content is greater than 48%, some glutenin will form gluten with great force, and some glutenin will form gluten with weak force. The gluten index is always about 92% in 64–100% HMW-GS relative content. The regression analysis indicated that SDS sediment value, development time and tolerance index were related linearly to the HMW-GS content, while the time to breakdown and become stable was exponentially relative.