[Crude extraction and activity assay of CEL I].
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.
Han
Suo-Yi
SY
Soybean Research Institute, Nanjing Agricultural University, Nanjing, China.
Yang
Ma-Li
ML
Gai
Jun-Yi
JY
Yu
De-Yue
DY
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
D003854:Deoxyribonucleotides; D010936:Plant Extracts; D043182:Carboxylesterase