Purification, Cloning, and Characterization of the CEL I Nuclease

Yang, Bing; Wen, Xiao; Kodali, Nagendra S.; Oleykowski, Catherine A.; Miller, C G; Kulinski, Joanne; Besack, David; Yeung, Jason A.; Kowalski, David; Yeung, Anthony T.

doi:10.1021/bi992376z

Cited by 115 publications

(96 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…There are several reports that EMC using CEL I nuclease demonstrates a very high sensitivity [70,73]. According to Qiu et al [71], the SURVEYOR Nuclease S corresponds to CEL II nuclease, a putative isozyme of CEL I nuclease [65]. EMC can be applied to longer PCR fragments compared to SSCP and HA [69,70].…”

Section: Discussionmentioning

confidence: 99%

“…Also, this EMC method is possible to apply to fluorescence automated sequencer [64,65,87] and CE platform [73] according to your laboratory equipments. We have established an ideal mutation screening system which is available to most any researcher.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, there are several reports of EMC using different kinds of enzymes [56][57][58][59][60][61][62][63]. Among them CEL nuclease is potentially the most effective enzyme for EMC reported recently [64][65][66][67][68][69][70] and is now commercially available as SURVEYOR Nuclease S from Transgenomic (Omaha, NE, USA) [71][72][73].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Development of a simple and highly sensitive mutation screening system by enzyme mismatch cleavage with optimized conditions for standard laboratories

Tsuji

Niida

2008

Electrophoresis

View full text Add to dashboard Cite

Efficient screening of unknown DNA variations is one of the substantive matters of molecular biology even today. Historically, single-strand conformation polymorphism (SSCP) and heteroduplex analysis (HA) are the most commonly used methods for detecting DNA variations everywhere in the world because of their simplicity. However, the sensitivity of these methods is not satisfactory for screening purpose. Recently, several new PCR based mutation screening methods have been developed, but most of them require special instruments and adjustment of conditions for each DNA sequence to attain the maximum sensitivity, eventually becoming as inconvenient as old methods. Enzyme mismatch cleavage (EMC) is potentially an ideal screening method. With high performance nucleases and once experimental conditions are optimized, it requires only conventional staff and conditions remain the same for each PCR product. In this study we tested four commercially available endonucleases for EMC and optimized the electrophoresis and developing conditions. We prepared 25 known DNA variations consisting of 18 single base substitutions (8 transitions and 10 transversions, including all possible sets of mismatches) and 7 small deletions or insertions. The combination of CEL nuclease, 12% polyacrylamide gel electrophoresis and rapid silver staining can detect all types of mutations and achieved 100% sensitivity.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Development of a simple and highly sensitive mutation screening system by enzyme mismatch cleavage with optimized conditions for standard laboratories

Tsuji

Niida

2008

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…The PCR reaction, using Pfu DNA polymerase, consisted of 40 cycles at 96°for 30 sec, 65°for 30 sec, and 72°for 1 min. CEL I was purified from celery stalks as described previously (Yang et al 2000).…”

Section: à2mentioning

confidence: 99%

Map-Based Cloning of the Gene Associated With the Soybean Maturity Locus E3

et al. 2009

View full text Add to dashboard Cite

Photosensitivity plays an essential role in the response of plants to their changing environments throughout their life cycle. In soybean [Glycine max (L.) Merrill], several associations between photosensitivity and maturity loci are known, but only limited information at the molecular level is available. The FT3 locus is one of the quantitative trait loci (QTL) for flowering time that corresponds to the maturity locus E3. To identify the gene responsible for this QTL, a map-based cloning strategy was undertaken. One phytochrome A gene (GmPhyA3) was considered a strong candidate for the FT3 locus. Allelism tests and gene sequence comparisons showed that alleles of Misuzudaizu (FT3/FT3; JP28856) and Harosoy (E3/E3; PI548573) were identical. The GmPhyA3 alleles of Moshidou Gong 503 (ft3/ft3; JP27603) and L62-667 (e3/e3; PI547716) showed weak or complete loss of function, respectively. High red/far-red (R/FR) long-day conditions enhanced the effects of the E3/FT3 alleles in various genetic backgrounds. Moreover, a mutant line harboring the nonfunctional GmPhyA3 flowered earlier than the original Bay (E3/E3; PI553043) under similar conditions. These results suggest that the variation in phytochrome A may contribute to the complex systems of soybean flowering response and geographic adaptation.

show abstract

“…CEL-I enzyme was isolated from celery according to Oleykowski et al (1998) and Yang et al (2000), with minor modifications (see our Web site for a detailed protocol, http://cuppen.niob. knaw.nl).…”

Section: Cel-i-mediated Library Screeningmentioning

confidence: 99%

Efficient Target-Selected Mutagenesis in Zebrafish

Wienholds

Eeden²,

Kosters³

et al. 2003

Genome Research

411

295

View full text Add to dashboard Cite

One of the most powerful methods available to assign function to a gene is to inactivate or knockout the gene. Recently,we described the first target-selected knockout in zebrafish. Here,we report on the further improvements of this procedure,resulting in a highly efficient and easy method to do target-selected mutagenesis in zebrafish. A library of 4608 ENU-mutagenized F1 animals was generated and kept as a living stock. The DNA of these animals was screened for mutations in 16 genes by use of CEL-I-mediated heteroduplex cleavage (TILLING) and subsequent resequencing. In total,255 mutations were identified,of which 14 resulted in a premature stop codon,7 in a splice donor/acceptor site mutation,and 119 in an amino acid change. By this method,we potentially knocked out 13 different genes in a few months time. Furthermore,we show that TILLING can be used to detect the full spectrum of ENU-induced mutations in a vertebrate genome with the presence of many naturally occurring polymorphisms

show abstract

Purification, Cloning, and Characterization of the CEL I Nuclease

Cited by 115 publications

References 22 publications

Development of a simple and highly sensitive mutation screening system by enzyme mismatch cleavage with optimized conditions for standard laboratories

Development of a simple and highly sensitive mutation screening system by enzyme mismatch cleavage with optimized conditions for standard laboratories

Map-Based Cloning of the Gene Associated With the Soybean Maturity Locus E3

Efficient Target-Selected Mutagenesis in Zebrafish

Contact Info

Product

Resources

About