Abstract: Background and Objective: Ginkgo biloba is a precious medicinal plant and has a long juvenile phase and spends 15-20 years in the vegetative phase before turning to reproductive phases, which makes breeding and cultivation of Ginkgo especially challenging. The FRI gene can regulate the FLC gene which inhibits flowering and further causes the late flowering of G. biloba. Therefore, the cloning and analysis of FRI gene can regulate the flowering time of G. biloba. Materials and Methods: The GbFRI gene and the protein sequence were analyzed using the online website of National Center for Biotechnology Information (NCBI), ProtParam and bioinformatic software of Clustal X2.0, Vector NTI 11.5 and MEGA6. The expression of GbFRI gene in different tissues of G. biloba was studied by quantitative RT-PCR (qRT-PCR). Data were analyzed with one-way ANOVA using SPSS11.0 for Windows. Results: The full length cDNA of GbFRI gene was 1702 bp (GenBank accession no. KY662058) and the open reading frame (ORF) covered 1602 bp, which encoded a 534 amino-acid protein. The predicted protein showed that a FRI superfamily and contain coiled-coil domains in two positions (between amino acids 55-100 and 405-450, respectively). The expression analysis results displayed that the highest GbFRI expression was in the male flowers. The GbFRI expression was higher in female flowers, stems than in the roots and fruits. The lowest relative expression of GbFRI was in the leaves. Conclusion: The GbFRI gene was isolated and characterized, laying a foundation for further study of vernalization pathway in G. biloba.