Detection of sequence heterology by use of the N. crassa nucleases

Bartok, Katalina; Fraser, Murray; Fareed, George C.

doi:10.1016/0006-291x(74)90269-1

Cited by 11 publications

(5 citation statements)

References 19 publications

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“…The N. crassa nuclease was employed for the isolation of pure lac operon [375], isolation of tRNA and rRNA gene hybrids [376,377] and for the detection of sequence heterology [378]. BAL 31 nuclease shows exonuclease activity and shortens duplex DNA from both 3′‐ and 5′‐ends.…”

Section: Applicationsmentioning

confidence: 99%

Single-strand-specific nucleases

2003

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Single-strand-specific nucleases are multifunctional enzymes and widespread in distribution. Their ability to act selectively on single-stranded nucleic acids and single-stranded regions in double-stranded nucleic acids has led to their extensive application as probes for the structural determination of nucleic acids. Intracellularly, they have been implicated in recombination, repair and replication, whereas extracellular enzymes have a role in nutrition. Although more than 30 single-strand-specific nucleases from various sources have been isolated till now, only a few enzymes (S1 nuclease from Aspergillus oryzae, P1 nuclease from Penicillium citrinum and nucleases from Alteromonas espejiana, Neurospora crassa, Ustilago maydis and mung bean) have been characterized to a significant extent. Recently, some of these enzymes have been cloned, their crystal structures solved and their interactions with different substrates have been established. The detection, purification, characteristics, structure-function correlations, biological role and applications of single-strand-specific nucleases are reviewed.

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Section: Applicationsmentioning

confidence: 99%

Single-strand-specific nucleases

2003

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“…Since their discovery, sugar non‐specific endonucleases have been extensively used as analytical tools for the determination of nucleic acid structure. The N. crassa nuclease was employed for the isolation of pure lac operon [232], isolation of tRNA and rRNA gene hybrids [233,234] and detection of sequence heterology [235]. Staphylococcal nuclease has been used to monitor hybridization reactions with DNA [14] and as a probe for drug binding sites on DNA [19].…”

Section: Applicationsmentioning

confidence: 99%

Sugar non-specific endonucleases

Rangarajan

Shankar

2001

FEMS Microbiol Rev

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Sugar non-specific endonucleases are multifunctional enzymes and are widespread in distribution. Apart from nutrition, they have also been implicated in cellular functions like replication, recombination and repair. Their ability to recognize different DNA structures has also been exploited for the determination of nucleic acid structure. Although more than 30 non-specific endonucleases have been isolated to date, very little information is available regarding their structure-function correlations except that of staphylococcal and Serratia nucleases. However, during the past few years, the primary structure, nature of the active site based on sequence homology, and the probable mechanism of action have been postulated for some of the enzymes. This review describes the purification, characteristics, biological role and applications of sugar non-specific endonucleases.

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“…Single‐strand specific nucleases such as S1, P1, SP, mung bean and CEL I nucleases have also been used to cleave heteroduplex DNA at the mismatch site (Ahmad et al. , 1975; Bartok et al. , 1974; Dodgson and Wells, 1977; Iwamatsu et al.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Arabidopsis thaliana mismatch specific endonucleases: application to mutation discovery by TILLING in pea

et al. 2007

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). † These authors contributed equally to this work. SummaryScanning DNA sequences for mutations and polymorphisms has become one of the most challenging, often expensive and time-consuming obstacles in many molecular genetic applications, including reverse genetic and clinical diagnostic applications. Enzymatic mutation detection methods are based on the cleavage of heteroduplex DNA at the mismatch sites. These methods are often limited by the availability of a mismatchspecific endonuclease, their sensitivity in detecting one allele in a pool of DNA and their costs. Here, we present detailed biochemical analysis of five Arabidopsis putative mismatch-specific endonucleases. One of them, ENDO1, is presented as the first endonuclease that recognizes and cleaves all types of mismatches with high efficiency. We report on a very simple protocol for the expression and purification of ENDO1. The ENDO1 system could be exploited in a wide range of mutation diagnostic tools. In particular, we report the use of ENDO1 for discovery of point mutations in the gibberellin 3b-hydrolase gene of Pisum sativum. Twenty-one independent mutants were isolated, five of these were characterized and two new mutations affecting internodes length were identified. To further evaluate the quality of the mutant population we screened for mutations in four other genes and identified 5-21 new alleles per target. Based on the frequency of the obtained alleles we concluded that the pea population described here would be suitable for use in a large reverse-genetics project.

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Detection of sequence heterology by use of the N. crassa nucleases

Cited by 11 publications

References 19 publications

Single-strand-specific nucleases

Single-strand-specific nucleases

Sugar non-specific endonucleases

Characterization of Arabidopsis thaliana mismatch specific endonucleases: application to mutation discovery by TILLING in pea

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