Purification, cloning, and characterization of the CEL I nuclease

B YangA T Yeung


CEL I, isolated from celery, is the first eukaryotic nuclease known that cleaves DNA with high specificity at sites of base-substitution mismatch and DNA distortion. The enzyme requires Mg(2+) and Zn(2+) for activity, with a pH optimum at neutral pH. We have purified CEL I 33 000-fold to apparent homogeneity. A key improvement is the use of alpha-methyl-mannoside in the purification buffers to overcome the aggregation of glycoproteins with endogenous lectins. The SDS gel electrophoresis band for the homogeneous CEL I, with and without the removal of its carbohydrate moieties, was extracted, renatured, and shown to have mismatch cutting specificity. After determination of the amino acid sequence of 28% of the CEL I polypeptide, we cloned the CEL I cDNA. Potential orthologs are nucleases putatively encoded by the genes BFN1 of Arabidopsis, ZEN1 of Zinnia, and DSA6 of daylily. Homologies of CEL I with S1 and P1 nucleases are much lower. We propose that CEL I exemplifies a new family of neutral pH optimum, magnesium-stimulated, mismatch duplex-recognizing nucleases, within the S1 superfamily.


Nov 28, 2012·Applied Microbiology and Biotechnology·Hiroaki MonTakahiro Kusakabe
Aug 13, 2013·Molecular Biotechnology·Nikolaos VogiatzakisEmmanouel Kanavakis
Oct 24, 2007·Molecular Biotechnology·Nikolaos VogiatzakisEmmanouel Kanavakis
May 3, 2012·Analytical Chemistry·Holger SchulzeTill T Bachmann
Apr 18, 2012·Journal of Agricultural and Food Chemistry·Chia-Yun KoJei-Fu Shaw
Jan 10, 2012·Nature Protocols·Neville E SanjanaFeng Zhang
May 14, 2004·Nucleic Acids Research·Bradley J TillSteven Henikoff
Dec 1, 2011·Nucleic Acids Research·Ishtiaq SaaemJingdong Tian
Apr 27, 2013·Plant & Cell Physiology·Krzysztof LesniewiczElzbieta Poreba
Dec 13, 2006·Molecular Plant-microbe Interactions : MPMI·Kurt H LamourHarold J G Meijer
Nov 14, 2003·Genome Research·Erno WienholdsEdwin Cuppen
Sep 25, 2003·Annual Review of Plant Biology·Steven Henikoff, Luca Comai
Aug 5, 2008·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Michael L NickersonLee E Moore
Nov 22, 2013·BMC Plant Biology·Núria Sánchez-Pons, Carlos M Vicient
Jun 5, 2007·BMC Biotechnology·Maxim PimkinAnthony T Yeung
Mar 19, 2013·Nanoscale Research Letters·Muniza ZahidSung Ha Park
Jul 2, 2011·Plant Methods·Wilma SabettaCinzia Montemurro
May 29, 2009·Genetics·Satoshi WatanabeKyuya Harada
Mar 19, 2014·Journal of Experimental Botany·Wataru Sakamoto, Tsuneaki Takami
Jan 3, 2012·Trends in Biotechnology·Siying MaJingdong Tian
Oct 12, 2010·Pediatric Neurology·Jacek PilchAleksander L Sieroń
May 8, 2010·Journal of Plant Physiology·Krzysztof LeśniewiczElzbieta Poreba
Jun 16, 2009·The Journal of Molecular Diagnostics : JMD·Konstantinos Voskarides, Constantinos Deltas
Oct 1, 2015·Journal of Biotechnology·Kun WangGeng Sheng Cao
Jan 26, 2007·The Journal of Molecular Diagnostics : JMD·Yanggu ShiGary F Gerard
Jul 27, 2007·The Plant Journal : for Cell and Molecular Biology·Karine TriquesAbdelhafid Bendahmane
Jan 31, 2006·The Plant Journal : for Cell and Molecular Biology·Luca Comai, Steven Henikoff
Feb 22, 2008·Electrophoresis·Michael J CrossRobert J Henry
May 14, 2011·Molecular Ecology Resources·M R GarvinA J Gharrett
Sep 3, 2013·Journal of Integrative Plant Biology·Aixia LiAimin Zhang
Mar 15, 2006·Biochemical and Biophysical Research Communications·Maxim PimkinAnthony T Yeung
Jan 13, 2006·Trends in Plant Science·Robbie WaughDavid Caldwell
May 11, 2005·Trends in Plant Science·Michael K UdvardiJens Stougaard

❮ Previous
Next ❯

Related Concepts

Trending Feeds


Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.


Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.


Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.