Inhibition of binding of phospholipase CΓ1 SH2 domains to phosphorylated epidermal growth factor receptor by phosphorylated peptides

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72540/1/j.1399-3011.1993.tb00138.x.pd

PerM: efficient mapping of short sequencing reads with periodic full sensitive spaced seeds

Motivation: The explosion of next-generation sequencing data has spawned the design of new algorithms and software tools to provide efficient mapping for different read lengths and sequencing technologies. In particular, ABI's sequencer (SOLiD system) poses a big computational challenge with its capacity to produce very large amounts of data, and its unique strategy of encoding sequence data into color signals

Landscape Features Impact on Soil Available Water, Corn Biomass, and Gene Expression during the Late Vegetative Stage

Crop yields at summit positions of rolling landscapes often are lower than backslope yields. The differences in plant response may be the result of many different factors. We examined corn (Zea mays L.) plant productivity, gene expression, soil water, and nutrient availability in two landscape positions located in historically high (backslope) and moderate (summit and shoulder) yielding zones to gain insight into plant response differences. Growth characteristics, gene expression, and soil parameters (water and N and P content) were determined at the V12 growth stage of corn. At tassel, plant biomass, N content, 13C isotope discrimination (Δ), and soil water was measured. Soil water was 35% lower in the summit and shoulder compared with the lower backslope plots. Plants at the summit had 16% less leaf area, biomass, and N and P uptake at V12 and 30% less biomass at tassel compared with plants from the lower backslope. Transcriptome analysis at V12 indicated that summit and shoulder-grown plants had 496 downregulated and 341 upregulated genes compared with backslope-grown plants. Gene set and subnetwork enrichment analyses indicated alterations in growth and circadian response and lowered nutrient uptake, wound recovery, pest resistance, and photosynthetic capacity in summit and shoulder-grown plants. Reducing plant populations, to lessen demands on available soil water, and applying pesticides, to limit biotic stress, may ameliorate negative water stress responses

Analysis of four PCR/SNaPshot multiplex assays analyzing 52 SNPforID markers

The SNPforID consortium identified a panel of 52 SNPs forensic analysis that has been used by several laboratories worldwide. The original analysis of the 52 SNPs was based on a single multiplex reaction followed by two single-base-extension (SBE) reactions each of which was analyzed using capillary electrophoresis. The SBE assays were designed for high throughput genetic analyzers and were difficult to use on the single capillary ABI PRISM 310 Genetic Analyzer and the latest generation 3500 Genetic Analyzer, as sensitivity on the 310 was low and separation of products on the 3500 with POP-7™ was poor. We have modified the original assay and split it into four multiplex reactions, each followed by an SBE assay. These multiplex assays were analyzed using polymer POP-4™ on ABI 310 PRISM® and polymers POP-4™, POP-6™ and POP-7™ on the 3500 Genetic Analyzer. The assays were sensitive and reproducible with input DNA as low as 60 pg using both the ABI 310 and 3500. In addition, we found that POP-6™ was most effective with the 3500, based on the parameters that we assessed, achieving better separation of the small SBE products; this conflicted with the recommended use of POP-7™ by the instrument manufacturer. To support the use of the SNP panel in casework in Malaysia we have created an allele frequency database from 325 individuals, representing the major population groups within Malaysia. Population and forensic parameters were estimated for all populations and its efficacy evaluated using 51 forensic samples from challenging casework

Quantitative analysis of ERG expression and its splice isoforms in formalin-fixed, paraffin-embedded prostate cancer samples: Association with seminal vesicle invasion and biochemical recurrence

© American Society for Clinical Pathology. Objectives: The proto-oncogene ETS-related gene (ERG) is consistently overexpressed in prostate cancer. Alternatively spliced isoforms of ERG have variable biological activities; inclusion of exon 11 (72 base pairs [bp]) is associated with aggressiveness and progression of disease. Exon 10 (81 bp) has also been shown to be alternatively spliced. Within this study, we assess whether ERG protein, messenger RNA (mRNA), and ERG splice isoform mRNA expression is altered as prostate cancer progresses. Methods: Detection of the TMPRSS2-ERG fusion was done using direct methods (reverse transcription polymerase chain reaction [PCR] and fluorescence in situ hybridization) and indirect methods for ERG mRNA and protein expression using quantitative PCR and immunohistochemistry, respectively. A linear equation method was used to quantitatively determine relative proportions of ERG variants (ERG72/Δ72, ERG81/Δ81) for each sample. Results: ERG mRNA and protein expression is increased in patients with advanced prostate cancer, with higher levels of ERG expression significantly associated with seminal vesicle invasion (stage pT3b) and biochemical recurrence. Genes involved in cell migration and invasiveness (matrix metalloproteinase 7, osteopontin, and septin 9) are increased in prostate cancers that overexpress ERG. In addition, there is a clear indication of increased retention of exons 10 and 11 in prostate cancer. Conclusions: Analysis of ERG and its variants may be valuable in determining prognosis and development of prostate cancer

Plant genotype influences aquatic-terrestrial ecosystem linkages through timing and composition of insect emergence

Terrestrial leaf litter provides aquatic insects with an energy source and habitat structure,and species differences in litter can influence aquatic insect emergence. Emerging insects also provide energy to riparian predators. We hypothesized that plant genetics would influence the composition and timing of emerging insect communities among individual genotypes of Populus angustifolia varying in litter traits. We also compared the composition and timing of emerging insect communities on litter from mixed genotypes of three cross types of a hybridizing cottonwood complex: P. angustifolia, P. fremontii, and their F1 hybrids. Using litter harvested from an experimental common garden, we measured emerging insect community composition, abundance, and production for 12 weeks in large litter packs affixed with emergence traps. Five major findings emerged. (1) In support of the genetic similarity hypothesis, we found that, among P. angustifolia tree genotypes, litter from more closely related genotypes had more similar litter thickness, nitrogen concentrations, decomposition rates, and emerging insect communities. (2) Genetic similarity was not correlated with other litter traits, although the litter fungal community was a strong predictor of emerging insect communities. (3) Litter decomposition rate, which was the strongest predictor of emerging aquatic insect communities, was influenced by litter thickness, litter N, and the litter fungal community. (4) In contrast to strong community composition differences among P. angustifolia genotypes, differences in community composition between P. fremontii and P. angustifolia were only marginally significant, and communities on F1 hybrids were indistinguishable from P. angustifolia despite genetic and litter trait differences. (5) Mixed litter packs muted the genetic effects observed in litter packs con- sisting of single genotypes. These results demonstrate that the genetic structure of riparian forests can affect the composition and timing of aquatic insect emergence. Because many riparian trees are clonal, including P. angustifolia, large clone size is likely to result in patches of genetically structured leaf litter that may influence the timing and composition of insect emergence within watersheds. Riparian restoration efforts incorporating different tree genotypes could also influence the biodiversity of emerging aquatic insects. Our work illustrates the importance of plant genes for community and ecosystem processes in riparian corridors

Maximizing signal-to-noise ratio in the random mutation capture assay

The ‘Random Mutation Capture’ assay allows for the sensitive quantitation of DNA mutations at extremely low mutation frequencies. This method is based on PCR detection of mutations that render the mutated target sequence resistant to restriction enzyme digestion. The original protocol prescribes an end-point dilution to about 0.1 mutant DNA molecules per PCR well, such that the mutation burden can be simply calculated by counting the number of amplified PCR wells. However, the statistical aspects associated with the single molecular nature of this protocol and several other molecular approaches relying on binary (on/off) output can significantly affect the quantification accuracy, and this issue has so far been ignored. The present work proposes a design of experiment (DoE) using statistical modeling and Monte Carlo simulations to obtain a statistically optimal sampling protocol, one that minimizes the coefficient of variance in the measurement estimates. Here, the DoE prescribed a dilution factor at about 1.6 mutant molecules per well. Theoretical results and experimental validation revealed an up to 10-fold improvement in the information obtained per PCR well, i.e. the optimal protocol achieves the same coefficient of variation using one-tenth the number of wells used in the original assay. Additionally, this optimization equally applies to any method that relies on binary detection of a small number of templates