Current Issues in Molecular Biology

796 KiB

Open AccessReview

The Ger Receptor Family from Sporulating Bacteria

by Christian Ross and Ernesto Abel-Santos

Curr. Issues Mol. Biol. 2010, 12(3), 147-158; https://doi.org/10.21775/cimb.012.147 - 16 Nov 2009

Cited by 2 | Viewed by 670

Bacterial spores are specialized cells that are exceptionally resistant to environmental stress. Spores convert back to actively growing cells, a process called germination, upon nutrient detection. The most common, initial step in the germination process is the recognition of small molecule germinants by germination (Ger) receptors. Ger receptors are inner-membrane heterocomplexes formed by three distinct protein products, the A-, B-, and C-subunits. In this review, we discuss and contrast published reports on representative Ger receptors from different Bacilli and Clostridia. We also present evidence for unrecognized germination pathways independent of Ger receptors. We further emphasize the function of L-alanine as a universal germinant. We also comment on biochemical aspects of germinant recognition and interaction between Ger receptor proteins. We propose that there are six general strategies used by Bacilli and Clostridia to integrate multiple germination signals. The use of different germinant recognition strategies results in germination response flexibility. Consequently, sporulating bacterial species that use the same biomolecules as germination signals can have different germination profiles. Finally, we discuss future directions for understanding the function of Ger receptors. Full article

668 KiB

Open AccessReview

p53-Based Anti-Cancer Therapies: An Empty Promise?

by Nikolas Desilet, Tessa N. Campbell and Francis Y.M. Choy

Curr. Issues Mol. Biol. 2010, 12(3), 143-146; https://doi.org/10.21775/cimb.012.143 - 16 Nov 2009

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Abstract

Since its discovery in 1979, p53 has become the focus of intensive cancer-based research in laboratories around the world. The p53 protein mediates critical cellular functions including the response to genotoxic stress, differentiation, senescence, and apoptosis, and has been shown to be mutated in a large proportion of human cancers. These observations led many to speculate that targeting the p53 pathway would result in the development of successful anti-cancer treatments. In spite of this, 30 years later, p53 has yet to fulfill this promise. However, new insights into small molecule combination therapies, microRNA regulation, structuring of clinical trials, and potential involvement in stem cell regulation may help p53 reach its potential. Full article

1140 KiB

Open AccessReview

Green Technologies for Room Temperature Nucleic Acid Storage

by Eunice Wan, Matthew Akana, Jennifer Pons, Justin Chen, Stacy Musone, Pui-Yan Kwok and Wilson Liao

Curr. Issues Mol. Biol. 2010, 12(3), 135-142; https://doi.org/10.21775/cimb.012.135 - 2 Oct 2009

Cited by 4 | Viewed by 1021

Abstract

Maintaining the long-term integrity of nucleic acids in the laboratory has traditionally required the use of freezers. However, novel nucleic acid stabilization technologies may allow for the storage of DNA and RNA at room temperature in a cost-effective, environmentally friendly manner. In this study, we evaluated two novel products for room temperature DNA storage: Biomatrica's DNA SampleMatrix technology and GenVault's GenTegra DNA technology. We compared the integrity and quality of DNA stored using these products against DNA stored in a -20° C freezer by performing downstream testing with short range PCR, long range PCR, DNA sequencing, and SNP microarrays. In addition, we tested Biomatrica's RNAstable product for its ability to preserve RNA at room temperature for use in a quantitative reverse transcription PCR assay. Full article

2288 KiB

Open AccessReview

SPUD qPCR Assay Confirms PREXCEL-Q Software's Ability to Avoid qPCR Inhibition

by J.M. Gallup, F.B. Sow, A. Van Geelen and M.R. Ackermann

Curr. Issues Mol. Biol. 2010, 12(3), 129-134; https://doi.org/10.21775/cimb.012.129 - 22 Sep 2009

Viewed by 645

Abstract

Real-time quantitative polymerase chain reaction is subject to inhibition by substances that co-purify with nucleic acids during isolation and preparation of samples. Such materials alter the activity of reverse transcriptase (RT) and thermostable DNA polymerase enzymes on which the assay depends. When removal of inhibitory substances by column or reagent-based methods fails or is incomplete, the remaining option of appropriately, precisely and differentially diluting samples and standards to non-inhibitory concentrations is often avoided due to the logistic problem it poses. To address this, we invented the PREXCEL-Q software program to automate the process of calculating the non-inhibitory dilutions for all samples and standards after a preliminary test plate has been performed on an experimental sample mixture. The SPUD assay was used to check for inhibition in each PREXCEL-Q-designed qPCR reaction. When SPUD amplicons or SPUD amplicon-containing plasmids were spiked equally into each qPCR reaction, all reactions demonstrated complete absence of qPCR inhibition. Reactions spiked with ~15,500 SPUD amplicons yielded a Cq of 27.39 +/- 0.28 (at ~80.8% efficiency), while reactions spiked with ~7,750 SPUD plasmids yielded a Cq of 23.82 +/- 0.15 (at ~97.85% efficiency). This work demonstrates that PREXCEL-Q sample and standard dilution calculations ensure avoidance of qPCR inhibition. Full article

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Curr. Issues Mol. Biol., Volume 12, Issue 3 (October 2010) – 4 articles

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