title: [Crude extraction and activity assay of CEL I].
abstract: CEL I, extracted from celery, is the first known eukaryotic nuclease that cleaves DNA with high specificity at sites of base-substitution mismatch and DNA distortion. It is a key enzyme for TILLING research. Here we reported a crude extraction method and activity assay of CEL I. Incision at mismatches of single nucleotide suggested that CEL I can effectively detect DNA at G-->A base substitution and the result can be obtained from an ABI377 Sequencer. Therefore, the extracted enzyme can be used in TILLING.
authors.affiliation: Soybean Research Institute, Nanjing Agricultural University, Nanjing, China.
authors.forename: Suo-Yi
authors.initials: SY
authors.lastname: Han
authors.forename: Ma-Li
authors.initials: ML
authors.lastname: Yang
authors.forename: Jun-Yi
authors.initials: JY
authors.lastname: Gai
authors.forename: De-Yue
authors.initials: DY
authors.lastname: Yu
mesh_terms: D000077102:Apium / Q000201:enzymology*; D020137:Base Pair Mismatch; D043182:Carboxylesterase / Q000302:isolation & purification* / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D018745:Genome, Plant; D010936:Plant Extracts; D017354:Point Mutation; D013379:Substrate Specificity
chemical_list: D003854:Deoxyribonucleotides; D010936:Plant Extracts; D043182:Carboxylesterase