Soybean is an economically-important food crop that contains about 40% of grain protein. The soybeans harvested from China are mainly used for food， and breeding for higher protein content is one of the main targets in soybean. Exploration of the genes related to soybean protein content and development of molecular markers are thus of great significance. In this study， a recombinant inbred line population was used by crossing a low protein soybean variety‘ ZH35’ （ female parent） with the high protein accession‘ Tokachi nagaha’ （ male parent） that was introduced from Japan. With the Bulked Segregant Analysis（ BSA） analysis， nine SSR markers that associated to protein content were identified on three chromosomes， including chromosome 19 where no QTL was previously reported. This QTL qPRO-19-1 was allocated using an inclusive complete interval mapping method（ ICIM-ADD） to the genetic interval of the molecular markers SSR_19_38 and SSR_19_59. This elite allele， which was derived from‘ Tokachi Nagaha’ ， could explain 7.81% and 14.87% of phenotypic variations in F2： 15and F2： 16 with the LOD values of 3.43 and 3.98， respectively. The physical interval of qPRO-19-1 expanded 385 kb that harbored 36 annotated genes. Collectively， the genetic mapping of the new QTL qPRO-19-1 laid the foundation for map-based cloning gene and molecular marker assisted breeding for high protein soybean varieties.