Quantitative assessment and evaluation of ecological parameters and biodiversity conservation are prime concerns for long-term conservation of rare and endangered species and their associated habitats in any ecological region. In this study, Gansu Province, a biodiversity hotspot, was chosen as the research area. We predicted the distribution patterns of suitable habitats for rare and endangered species. The replacement cost method was adopted to calculate the conservation value of rare and endangered species. The suitable habitat distribution area of rare and endangered wild animals reached 351,607.76 km2 (without overlapping area), while that of plants reached 72,988.12 km2 (without overlapping area). The conservation value of rare and endangered wildlife is US $1670.00 million. The high-value areas are mostly concentrated in the south and north of Gansu Province. The conservation value of rare and endangered wild plants is US $56,920.00 million. The high-value areas are mostly concentrated south of Gansu Province. The conservation value is US $58,590.00 million a year, and its distribution trend is gradually decreasing from northeast to southwest, with the highest in the forest area south of Gansu Province, followed by the Qilian Mountain area in the north. These results are of great significance for future improvement of the evaluation index system of ecosystem services and the development of ecosystem services and management strategies.