Immobilized Nucleases

Reddy, Laxma G.; Shankar, V.

doi:10.3109/07388559309041320

Cited by 35 publications

(16 citation statements)

References 63 publications

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“…Their application depends on the specificity and mode of action of a particular enzyme. Nucleases have been employed in the production of flavor enhancers, removal of nucleic acids, as therapeutic agents, and for the determination of nucleic acid structure, in particular, mutation typing (Reddy and Shankar 1993;Gite and Shankar 1995;Jasin 1996;Singwi and Joshi 2000;Williams 2001). Nucleases with novel and unusual properties may facilitate the development of advanced technologies in many areas of biotechnology and biomedicine.…”

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confidence: 99%

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

Shagin¹,

Rebrikov²,

Kozhemyako³

et al. 2002

Genome Res.

229

202

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We have characterized a novel nuclease from the Kamchatka crab, designated duplex-specific nuclease (DSN). DSN displays a strong preference for cleaving double-stranded DNA and DNA in DNA-RNA hybrid duplexes, compared to single-stranded DNA. Moreover, the cleavage rate of short, perfectly matched DNA duplexes by this enzyme is essentially higher than that for nonperfectly matched duplexes of the same length. Thus, DSN differentiates between one-nucleotide variations in DNA. We developed a novel assay for single nucleotide polymorphism (SNP) detection based on this unique property, termed "duplex-specific nuclease preference" (DSNP). In this innovative assay, the DNA region containing the SNP site is amplified and the PCR product mixed with signal probes (FRET-labeled short sequence-specific oligonucleotides) and DSN. During incubation, only perfectly matched duplexes between the DNA template and signal probe are cleaved by DSN to generate sequence-specific fluorescence. The use of FRET-labeled signal probes coupled with the specificity of DSN presents a simple and efficient method for detecting SNPs. We have employed the DSNP assay for the typing of SNPs in methyltetrahydrofolate reductase, prothrombin and p53 genes on homozygous and heterozygous genomic DNA.

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confidence: 99%

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

Shagin¹,

Rebrikov²,

Kozhemyako³

et al. 2002

Genome Res.

229

202

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“…A general and effective technique to add and then eliminate an enzyme is by attaching it covalently to a solid support (6). The enzymatic side chains that are most amenable to covalent modification are cysteine, lysine, and histidine.…”

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confidence: 99%

A Highly Active Immobilized Ribonuclease

Sweeney

Kelemen

Woycechowsky

et al. 2000

Analytical Biochemistry

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“…Nucleic acid content of the SCP has to be decreased to provide the safe level (2 g per day) for human consumption. Enzymatic treatment was proposed as an effective method for removal of RNA from SCP without changing the protein content (Castro et al 1971;Yang et al 1979;Reddy and Shankar 1993). While Castro et al (1971) used bovine pancreatic RNase A for the reduction of nucleic acid content in Candida yeast; Yang et al (1979) presented a microbial source of RNase for reduction of endogenous nucleic acids in SCP as well as inclusion of functional groups for adding flavors.…”

Section: Use Of Microbial Rnases In Downstream Processesmentioning

confidence: 97%

“…In this regard, their intrinsic properties provide a therapeutic opportunity for treatment of cancer (Makarov and Ilinskaya 2003). On the other hand, RNases have also application potential for sequence and structure analysis of RNA, preparation of oligonucleotides, removal of RNA from single-cell protein (Reddy and Shankar 1993;Castro et al 1971;Yang et al 1979), preparation of pure plasmid DNA for DNA vaccines and gene therapy (Voss et al 2006;Han et al 2009). …”

Section: Introductionmentioning

confidence: 97%

Microbial ribonucleases (RNases): production and application potential

Hameş

Demir

2015

World J Microbiol Biotechnol

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Ribonuclease (RNase) is hydrolytic enzyme that catalyzes the cleavage of phosphodiester bonds in RNA. RNases play an important role in the metabolism of cellular RNAs, such as mRNA and rRNA or tRNA maturation. Besides their cellular roles, RNases possess biological activity, cell stimulating properties, cytotoxicity and genotoxicity. Cytotoxic effect of particular microbial RNases was comparable to that of animal derived counterparts. In this respect, microbial RNases have a therapeutic potential as anti-tumor drugs. The significant development of DNA vaccines and the progress of gene therapy trials increased the need for RNases in downstream processes. In addition, RNases are used in different fields, such as food industry for single cell protein preparations, and in some molecular biological studies for the synthesis of specific nucleotides, identifying RNA metabolism and the relationship between protein structure and function. In some cases, the use of bovine or other animal-derived RNases have increased the difficulties due to the safety and regulatory issues. Microbial RNases have promising potential mainly for pharmaceutical purposes as well as downstream processing. Therefore, an effort has been given to determination of optimum fermentation conditions to maximize RNase production from different bacterial and fungal producers. Also immobilization or strain development experiments have been carried out.

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Immobilized Nucleases

Cited by 35 publications

References 63 publications

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

A Highly Active Immobilized Ribonuclease

Microbial ribonucleases (RNases): production and application potential

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