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31.
Background
Statistical methods for identifying positively selected sites in protein coding regions are one of the most commonly used tools in evolutionary bioinformatics. However, they have been limited by not taking the physiochemical properties of amino acids into account. 相似文献32.
M. García-Closas L. E. Moore C. S. Rabkin T. Franklin J. Struewing D. Ginzinger 《Biomarkers》2013,18(5):472-479
AbstractBuccal cell samples are increasingly used in epidemiological studies as a source of genomic DNA. The accurate and precise quantitation of human DNA is critical for the optimal use of these samples. However, it is complicated by the presence of bacterial DNA and wide inter-individual variation in DNA concentration from buccal cell collections. The paper evaluated the use of ultraviolet light (UV) spectroscopy, Höechst (H33258) and PicoGreen? as measures of total DNA, and real-time quantitative polymerase chain reaction (PCR) as a measure of human amplifiable DNA in buccal samples. Using serially diluted white blood cell DNA samples (at a concentration range of 300 to 0.5?ng µl?1), UV spectroscopy showed the largest bias, followed by Höechst, especially for low concentrations. PicoGreen and real-time PCR provided the most accurate and precise estimates across the range of concentrations evaluated, although an increase in bias with decreasing concentrations was observed. The ratio of real-time PCR to PicoGreen provided a reasonable estimate of the percentage of human DNA in samples containing known mixtures of human and bacterial DNA. Quantification of buccal DNA from samples collected in a breast cancer case-control study by PicoGreen and real-time PCR indicated that cytobrush and mouthwash DNA samples contain similar percentages of human amplifiable DNA. Real-time PCR is recommended for the quantification of buccal cell DNA in epidemiological studies since it provides precise estimates of human amplifiable DNA across the wide range of DNA concentrations commonly observed in buccal cell DNA samples. 相似文献
33.
A new computer program, called SHIFTX2, is described which is capable of rapidly and accurately calculating diamagnetic 1H, 13C and 15N chemical shifts from protein coordinate data. Compared to its predecessor (SHIFTX) and to other existing protein chemical
shift prediction programs, SHIFTX2 is substantially more accurate (up to 26% better by correlation coefficient with an RMS
error that is up to 3.3× smaller) than the next best performing program. It also provides significantly more coverage (up
to 10% more), is significantly faster (up to 8.5×) and capable of calculating a wider variety of backbone and side chain chemical
shifts (up to 6×) than many other shift predictors. In particular, SHIFTX2 is able to attain correlation coefficients between
experimentally observed and predicted backbone chemical shifts of 0.9800 (15N), 0.9959 (13Cα), 0.9992 (13Cβ), 0.9676 (13C′), 0.9714 (1HN), 0.9744 (1Hα) and RMS errors of 1.1169, 0.4412, 0.5163, 0.5330, 0.1711, and 0.1231 ppm, respectively. The correlation between SHIFTX2’s
predicted and observed side chain chemical shifts is 0.9787 (13C) and 0.9482 (1H) with RMS errors of 0.9754 and 0.1723 ppm, respectively. SHIFTX2 is able to achieve such a high level of accuracy by using
a large, high quality database of training proteins (>190), by utilizing advanced machine learning techniques, by incorporating
many more features (χ2 and χ3 angles, solvent accessibility, H-bond geometry, pH, temperature), and by combining sequence-based with structure-based chemical
shift prediction techniques. With this substantial improvement in accuracy we believe that SHIFTX2 will open the door to many
long-anticipated applications of chemical shift prediction to protein structure determination, refinement and validation.
SHIFTX2 is available both as a standalone program and as a web server (). 相似文献
34.
Ginzinger SW Gruber M Brandstetter H Sippl MJ 《Structure (London, England : 1993)》2011,19(12):1739-1743
Recurring groups of atoms in molecules are surrounded by specific canonical distributions of electrons. Deviations from these distributions reveal unrealistic molecular geometries. Here, we show how canonical electron densities can be combined with classical electron densities derived from X-ray diffraction experiments to drive the real space refinement of crystal structures. The refinement process generally yields superior molecular models with reduced excess electron densities and improved stereochemistry without compromising the agreement between molecular models and experimental data. 相似文献
35.
D I Smith W Liu D Ginzinger P Green S Smith N D Wang F Recchia K Carolyn H Drabkin W Golembieski 《Genomics》1991,11(1):179-187
A total of 5700 human chromosome 3-specific cosmid clones was isolated from a series of cosmid libraries constructed from somatic cell hybrids whose only human component was an entire chromosome 3 or a chromosome 3 containing an interstitial deletion removing 50% of long arm sequences. Several unique sequence chromosome 3-specific hybridization probes were isolated from each of 616 of these cosmids. These probes were then used to localize the cosmids by hybridization to a somatic cell hybrid deletion mapping panel capable of resolving chromosome 3 into nine distinct subregions. All 616 of the cosmids were localized to either the long or short arm of chromosome 3 and 63% of the short arm cosmids were more precisely localized. We have identified a total of 87 cosmids that contain fragments that are evolutionarily conserved. Fragments from these cosmids should prove useful in the identification of new chromosome 3-specific genes as well as in comparative mapping studies. The localized cosmids should provide excellent saturation of human chromosome 3 and facilitate the construction of physical and genetic linkage maps to identify various disease loci including Von Hippel Lindau disease and renal and small cell lung carcinoma. 相似文献
36.
Background
Alternative exons encode different isoforms of the human insulin-like growth factor-I (IGF-I) precursor without altering mature IGF-I. We hypothesized that the various IGF-I precursors may traffic IGF-I differently. Chimeric IGF-I precursors were made with green fluorescent protein (GFP) cloned between the signal and mature IGF-I domains. 相似文献37.
The construction of a consistent protein chemical shift database is an important step toward making more extensive use of
this data in structural studies. Unfortunately, progress in this direction has been hampered by the quality of the available
data, particularly with respect to chemical shift referencing, which is often either inaccurate or inconsistently annotated.
Preprocessing of the data is therefore required to detect and correct referencing errors. We have developed a program for
performing this task, based on the comparison of reported and expected chemical shift distributions. This program, named CheckShift,
does not require additional data and is therefore applicable to data sets where structures are not available. Therefore CheckShift
provides the possibility to re-reference chemical shifts prior to their use as structural constraints. 相似文献
38.
Autologous disc cell implantation, growth factors and gene therapy appear to be promising therapies for disc regeneration. Unfortunately, the replicative lifespan and growth kinetics of human nucleus pulposus (NP) cells related to host age are unclear. We investigated the potential relations among age, replicative lifespan and growth rate of NP cells, and determined the age range that is suitable for cell-based biological therapies for degenerative disc diseases. We used NP tissues classified by decade into five age groups: 30s, 40s, 50s, 60s and 70s. The mean cumulative population doubling level (PDL) and population doubling rate (PDR) of NP cells were assessed by decade. We also investigated correlations between cumulative PDL and age, and between PDR and age. The mean cumulative PDL and PDR decreased significantly in patients in their 60s. The mean cumulative PDL and PDR in the younger groups (30s, 40s and 50s) were significantly higher than those in the older groups (60s and 70s). There also were significant negative correlations between cumulative PDL and age, and between PDR and age. We found that the replicative lifespan and growth rate of human NP cells decreased with age. The replicative potential of NP cells decreased significantly in patients 60 years old and older. Young individuals less than 60 years old may be suitable candidates for NP cell-based biological therapies for treating degenerative disc diseases. 相似文献
39.
Ilke van Hazel Francesco Santini Johannes Müller Belinda SW Chang 《BMC evolutionary biology》2006,6(1):97-15
Background
Vertebrate SWS1 visual pigments mediate visual transduction in response to light at short wavelengths. Due to their importance in vision, SWS1 genes have been isolated from a surprisingly wide range of vertebrates, including lampreys, teleosts, amphibians, reptiles, birds, and mammals. The SWS1 genes exhibit many of the characteristics of genes typically targeted for phylogenetic analyses. This study investigates both the utility of SWS1 as a marker for inferring vertebrate phylogenetic relationships, and the characteristics of the gene that contribute to its phylogenetic utility. 相似文献40.