A method for homologous recombination comprising: using a PCR product and a linearized vector, wherein the PCR product contains a target gene sequence having amplification primer sequences P1 and P2 on its both ends, and the linearized vector has homologous recombination regions VP1 and VP2 comprising nucleotide sequences homologous to the amplification primer sequences P1 and P2 of the PCR product and the linearized vector also has a homologous recombination region VT1 comprising a nucleotide sequence homologous to a sequence T1 which is a part of sequence in the PCR product internal to P1 on the terminal side of VP1 and/or a homologous recombination region VT2 comprising a nucleotide sequence homologous to a sequence T2 which is a part of sequence in the PCR product internal to P2 on the terminal side of VP2, provided that at least one of T1 and T2 has a nucleotide sequence specific to the target gene; wherein the PCR product is prepared by two times of PCR operations, the first PCR operation is conducted by using an amplification primer containing T1 sequence and an amplification primer containing P3 sequence, provided that the P3 sequence has no homology to any of homologous recombination regions of the vector VT1, VP1, VT2 and VP2, and the second PCR operation is conducted by using an amplification primer containing P1 sequence and an amplification primer containing P2 sequence; wherein the 3'-terminal side region of the amplification primer containing P1 sequence and the 5'-terminal side region of the amplification primer containing T1 sequence have a partially overlapped sequence, and the amplification primer containing P2 sequence and the amplification primer containing P3 sequence have a partially overlapped sequence or P2 sequence is internal to P3 sequence; making the PCR product be subject to homologous recombination reaction and thereby be inserted into the vector, wherein the homologous recombination reaction is conducted in a cell by using Red (Red α/β) or RecE/T system or is conducted by using In-Fusion® method; and obtaining a recombinant DNA molecule in which a PCR product of interest is specifically inserted into a vector. using a PCR product and a linearized vector, wherein the PCR product contains a target gene sequence having amplification primer sequences P1 and P2 on its both ends, and the linearized vector has homologous recombination regions VP1 and VP2 comprising nucleotide sequences homologous to the amplification primer sequences P1 and P2 of the PCR product and the linearized vector also has a homologous recombination region VT1 comprising a nucleotide sequence homologous to a sequence T1 which is a part of sequence in the PCR product internal to P1 on the terminal side of VP1 and/or a homologous recombination region VT2 comprising a nucleotide sequence homologous to a sequence T2 which is a part of sequence in the PCR product internal to P2 on the terminal side of VP2, provided that at least one of T1 and T2 has a nucleotide sequence specific to the target gene; wherein the PCR product is prepared by two times of PCR operations, the first PCR operation is conducted by using an amplification primer containing T1 sequence and an amplification primer containing P3 sequence, provided that the P3 sequence has no homology to any of homologous recombination regions of the vector VT1, VP1, VT2 and VP2, and the second PCR operation is conducted by using an amplification primer containing P1 sequence and an amplification primer containing P2 sequence; wherein the 3'-terminal side region of the amplification primer containing P1 sequence and the 5'-terminal side region of the amplification primer containing T1 sequence have a partially overlapped sequence, and the amplification primer containing P2 sequence and the amplification primer containing P3 sequence have a partially overlapped sequence or P2 sequence is internal to P3 sequence; making the PCR product be subject to homologous recombination reaction and thereby be inserted into the vector, wherein the homologous recombination reaction is conducted in a cell by using Red (Red α/β) or RecE/T system or is conducted by using In-Fusion® method; and obtaining a recombinant DNA molecule in which a PCR product of interest is specifically inserted into a vector.