title: Molecular cloning, expression, and characterization of a Ca2+-dependent nuclease of Arabidopsis thaliana.
abstract: A gene encoding a Ca(2+)-dependent nuclease (AtCaN2) consisting of 226 amino acids from Arabidopsis thaliana was isolated. The AtCaN2 gene was expressed in Escherichia coli as a His fusion protein. The His-AtCaN2 fusion protein was purified by affinity chromatography using a Ni-NTA column. Western blot analysis using an anti-His antibody showed that the His-AtCaN2 fusion protein was not degraded. The nuclease activity of the His-AtCaN2 fusion protein was highest at 37°C, required a neutral or weakly-alkaline environment, and was stimulated by Ca(2+) and Mg(2+), but inhibited by Zn(2+) and high concentration of Mn(2+).
authors.affiliation: Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin, China.
authors.forename: Kunyuan
authors.initials: K
authors.lastname: Guo
authors.forename: Shenkui
authors.initials: S
authors.lastname: Liu
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Takano
authors.forename: Xinxin
authors.initials: X
authors.lastname: Zhang
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D004720:Endonucleases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D011993:Recombinant Fusion Proteins; D004720:Endonucleases; D002118:Calcium