A Ni(Ⅱ) coordination complex ［Ni(2,2′-biby) (H2O)4］·(tba)2·5H2O(2,2′-biby=2,2′-dipyridine, H2tba=2-thiobarbituric acid ) has been synthesized and structurally characterized by IR, element analysis and single crystal X-ray diffractometry. The results of single crystal X-ray diffractometry displayed that the title compound belongs to orthorhombic with space group of Pbca, and its cell parameters are a=13.662(2) , b=19.470(4) , c=21.590(4)  . The binding of title compound with Herring sperm DNA/BSA has been investigated by absorption and fluorescence spectra, and the results displayed that the title compound binds with DNA in electrostatic interaction mode, and quenches the intrinsic fluorescence of BSA by a static quenching mechanism.