Ribosomal protein L6 (RpL6, also called Taxreb107) possesses at least three nuclear localization signal (NLS)-like motifs. The activity of these motifs for their ability to mediate protein nuclear translocation was analyzed by using a NLS trapping system established. The full length or different fragments of RpL6/Taxreb107 cDNA was inserted to the cloning site of NLS trapping vector and the resulting constructs were used for transformation of host yeast. The result showed that the first two NLS-like motifs of RpL6/Taxreb107 induced the fusion protein to be transfected into the nucleus while the third one did not. This conclusion was confirmed by transfection of cultured cells with (EGFP) fused with different RpL6/Taxreb107 fragments. The results also showed that the first two NLS-like motifs of RpL6/Taxreb107 have nucleolus localization activity. When expressed in cultured cells, the RpL6/Taxreb107 fragments containing the first two NLS preferentially induced the fusion protein to be transfected into the nucleoli. These results are helpful for understanding of the nuclear translocation of RpL6/Taxreb107, and also confirmed that the NLS-trapping system is useful for searching NLS in proteins.