Bimolecular fluorescence complementation technology capable of effectively identifying false positive signal

The invention discloses a bimolecular fluorescent complementation (BiFC) technology capable of effectively identifying a false positive signal. The existing BiFC imaging technology is improved by a method of adding reference fluorescent protein. The reference fluorescent protein is connected with an N-terminal or C-terminal fragment of a labeled fluorescent protein to form a fusion protein, the expression level of a plasmid vector is represented through the fluorescence signal intensity of the reference fluorescent protein, the fluorescence signal intensity of the labeled fluorescent protein representing a BiFC signal is subjected to normalization processing, and the fluorescence signal intensity ratio of the labeled fluorescent protein to the reference fluorescent protein is used as a parameter for identifying the false positive signal. The technology can be easily and efficiently applied to various BiFC marking systems for researching the protein-protein interaction, the false positive signal is identified