Current Issues in Molecular Biology

539 KiB

Open AccessReview

PNA Biosensors for Nucleic Acid Detection

by Joseph Wang

Curr. Issues Mol. Biol. 1999, 1(2), 117-122; https://doi.org/10.21775/cimb.001.117 - 31 Jul 1999

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Biosensor devices, based on the conversion of nucleic acid recognition reactions into useful electrical signals, offer considerable promise for DNA diagnostics. The unique hybridization properties of solution-phase PNA can be extrapolated onto transducer surfaces in connection with the design of remarkably specific DNA [...] Read more.

Biosensor devices, based on the conversion of nucleic acid recognition reactions into useful electrical signals, offer considerable promise for DNA diagnostics. The unique hybridization properties of solution-phase PNA can be extrapolated onto transducer surfaces in connection with the design of remarkably specific DNA biosensors. This article reviews the development of PNA biosensors, and discusses common PNA-biosensing protocols along with their prospects in DNA biosensor technology. Full article

989 KiB

Open AccessReview

Peptide Nucleic Acid (PNA) Antisense Effects in Escherichia coli

by Liam Good and Peter E. Nielsen

Curr. Issues Mol. Biol. 1999, 1(2), 111-116; https://doi.org/10.21775/cimb.001.111 - 31 Jul 1999

Cited by 2 | Viewed by 457

Abstract

Antisense peptide nucleic acid (PNA) can be used to control cell growth, gene expression and growth phenotypes in the bacteria Escherichia coli . PNAs targeted to the RNA components of the ribosome can inhibit translation and cell growth, and PNAs targeted to mRNA [...] Read more.

Antisense peptide nucleic acid (PNA) can be used to control cell growth, gene expression and growth phenotypes in the bacteria Escherichia coli . PNAs targeted to the RNA components of the ribosome can inhibit translation and cell growth, and PNAs targeted to mRNA can limit gene expression with gene and sequence specificity. In an E. coli cell extract, efficient inhibition is observed when using PNA concentrations in the nanomolar range, whereas micromolar concentrations are required for inhibition in growing cells. A mutant strain of E. coli that is more permeable to antibiotics also is more susceptible to antisense PNAs than the wild type. This chapter details methods for testing the antisense activities of PNA in E. coli. As an example of the specific antisense inhibition possible, we show the effects of an anti- ß-galactosidase PNA in comparison to control PNAs. With improvements in cell uptake, antisense PNAs may find applications as antimicrobial agents and as tools for microbial functional genomics. Full article

535 KiB

Open AccessReview

Purification of Nucleic Acids by Hybridization to Affinity Tagged PNA Probes

by Henrik Ørum

Curr. Issues Mol. Biol. 1999, 1(2), 105-110; https://doi.org/10.21775/cimb.001.105 - 31 Jul 1999

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Abstract

The use of affinity tagged PNA capture probes offers an efficient means for the purification of nucleic acids by hybridization. Two different approaches are described. A sequence specific method and a generic method. The sequence specific method requires sequence information on the target [...] Read more.

The use of affinity tagged PNA capture probes offers an efficient means for the purification of nucleic acids by hybridization. Two different approaches are described. A sequence specific method and a generic method. The sequence specific method requires sequence information on the target and synthesis of a dedicated PNA. It can be used to selectively purify the nucleic acid containing the target from non-related nucleic acids and other cellular components. The generic method uses a "universal" triplex forming PNA and requires no sequence information on the target. It can be used in the bulk purification of large nucleic acids. Full article

774 KiB

Open AccessReview

An Introduction to Peptide Nucleic Acid

by Peter E. Nielsen and Michael Egholm

Curr. Issues Mol. Biol. 1999, 1(2), 89-104; https://doi.org/10.21775/cimb.001.089 - 31 Jul 1999

Cited by 5 | Viewed by 1805

Abstract

Peptide Nucleic Acid (PNA) is a powerful new biomolecular tool with a wide range of important applications. PNA mimics the behaviour of DNA and binds complementary nucleic acid strands. The unique chemical, physical and biological properties of PNA have been exploited to produce [...] Read more.

Peptide Nucleic Acid (PNA) is a powerful new biomolecular tool with a wide range of important applications. PNA mimics the behaviour of DNA and binds complementary nucleic acid strands. The unique chemical, physical and biological properties of PNA have been exploited to produce powerful biomolecular tools, antisense and antigene agents, molecular probes and biosensors. Full article

778 KiB

Open AccessReview

Endonuclease-Mediated Long PCR and Its Application to Restriction Mapping

by Chengtao Her and Richard M. Weinshilboum

Curr. Issues Mol. Biol. 1999, 1(2), 77-88; https://doi.org/10.21775/cimb.001.077 - 31 Jul 1999

Viewed by 410

Abstract

The polymerase chain reaction (PCR) is the most widely used technique for the study of DNA. Applications for PCR have been extended significantly by the development of "long" PCR, a technique that makes it possible to amplify DNA fragments up to 40 kb [...] Read more.

The polymerase chain reaction (PCR) is the most widely used technique for the study of DNA. Applications for PCR have been extended significantly by the development of "long" PCR, a technique that makes it possible to amplify DNA fragments up to 40 kb in length. This article describes two novel applications of the long PCR technique, one which simplifies restriction mapping and another which enhances amplification specificity and yield. The same primers used to perform the long PCR amplification can be used as probes to perform restriction mapping of the DNA fragment amplified. Restriction digestion performed prior to long PCR amplification can be used to selectively suppress the amplification of members of families of closely related DNA sequences, thereby making it possible to selectively amplify one of a group of highly homologous sequences. These two complimentary techniques, both involving use of the long PCR paired with restriction digestion, have potential application in any laboratory in which PCR is performed. Full article

1469 KiB

Open AccessReview

Cell Biological Studies of the Prion Protein

by David A. Harris

Curr. Issues Mol. Biol. 1999, 1(2), 65-76; https://doi.org/10.21775/cimb.001.065 - 31 Jul 1999

Viewed by 421

Abstract

Studying PrP^C and PrP^Sc in cell culture systems is advantageous because such systems contain all the organelles, membranes, and molecular cofactors that are likely to play an important role in the biology of the proteins. Using cultured cells expressing PrP^C [...] Read more.

Studying PrP^C and PrP^Sc in cell culture systems is advantageous because such systems contain all the organelles, membranes, and molecular cofactors that are likely to play an important role in the biology of the proteins. Using cultured cells expressing PrP^C, we have discovered that this isoform constitutively cycles between the cell surface and an endocytic compartment, a process that is mediated by clathrin-coated pits and a putative PrP^C receptor. We have also constructed stably transfected lines of CHO cells that express PrP molecules carrying mutations that are associated with familial prion diseases. The mutant PrP molecules in these cells are spontaneously converted to the PrP^Sc state, a phenomenon which has allowed us to analyze several key features of prion formation. Full article

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Curr. Issues Mol. Biol., Volume 1, Issue 2 (July 1999) – 6 articles

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