The disclosure discloses a preparation of type I collagen-like fibers and a method for regulating and controlling the periodic length of fiber stripes thereof, belonging to the technical field of genetic engineering. The disclosure produces a three-segment chimeric collagen P-CL-P pattern by inserting a continuous Gly-Xaa-Yaa triplet collagen sequence in the middle based on the N- and C-terminal (GPP)n sequences. The self-assembly is driven by the interaction between the N- and C-terminal (GPP)n triple helixes to form banded fibers with periodic bright and dark stripes. According to the method of the disclosure, a fiber from a clean source, which can self-assemble to form periodic bright and dark stripes can be prepared, the structure of which is similar to type I collagen, the preparation process is simple, the collagen fiber with low cost can be produced on a large scale, and the method has broad application prospects in the field of biological materials.