The invention belongs to the field of medical biotechnology, and relates to two series major vault protein of glucagon-like peptide-2 (GLP-2) of people and a preparation method of the two series major vault protein. A second place of an amino acid fragment of the glucagon-like peptide-2 of people is mutated, two segments are connected in series through connecting peptide, a dimmer structure of the GLP-2 is simulated, encoding gene of the two series major vault protein of the GLP-2 is obtained by a synthetic method, the encoding gene is cloned to a pET22b(+) pronucleus so as to express a vector, Escherichia coli BL21 is transformed, the encoding gene is expressed in the Escherichia coli efficiently, the purified reconstructed purpose protein is obtained through salt fractionation of ammonium sulfate in fractional and chromatography of anion exchange columns. The reconstructed purpose protein can stimulate a Caco-2 cell, multiplication of the Caco-2 cell can be promoted notably, the two series major vault protein of GLP-2 of people has natural biological activities, and therefore a foundation is established for further study and extensive use of the GLP-2 of people.