Projection structure of full length connexin 43 by electron cryo-crystallography

We previously used electron cryo-crystallography to determine the three-dimensional structure of recombinant gap junction channels formed by a C-terminal truncation mutant of Cx43 (11). The dodecameric channel is formed by the end-to-end docking of two hexameric connexons, each comprised of 24 transmembrane alpha-helices. We have now generated two-dimensional crystals of the recombinant, full-length channel, as well as crystals in which the C-tail has been completely removed by trypsin digestion. Projection density maps at 7.5 A resolution closely resemble our previous analysis of the C-terminal truncation mutant (9). A difference map between the full length and trypsin-treated channels suggests that there are small but significant shifts in protein density upon removal of the C-tail.     

Differences in Crossover Frequency and Distribution among Three Sibling Species of Drosophila

Comparisons of the genetic and cytogenetic maps of three sibling species of Drosophila reveal marked differences in the frequency and cumulative distribution of crossovers during meiosis. The maps for two of these species, Drosophila melanogaster and D. simulans, have previously been described, while this report presents new map data for D. mauritiana, obtained using a set of P element markers. A genetic map covering nearly the entire genome was constructed by estimating the recombination fraction for each pair of adjacent inserts. The P-based genetic map of mauritiana is ~1.8 times longer than the standard melanogaster map. It appears that mauritiana has higher recombination along the entire length of each chromosome, but the difference is greatest in centromere-proximal regions of the autosomes. The mauritiana autosomes show little or no centromeric recombinational suppression, a characteristic that is prominent in melanogaster. D. simulans appears to be intermediate both in terms of total map length and intensity of the autosomal centromeric effect. These interspecific differences in recombination have important evolutionary implications for DNA sequence organization and variability. In particular, mauritiana is expected to differ from melanogaster in patterns and amounts of sequence variation and transposon insertions.     

Projection Structure of Full Length Connexin 43 by Electron Cryo-Crystallography

We previously used electron cryo-crystallography to determine the three-dimensional structure of recombinant gap junction channels formed by a C-terminal truncation mutant of Cx43 (11). The dodecameric channel is formed by the end-to-end docking of two hexameric connexons, each comprised of 24 transmembrane α-helices. We have now generated two-dimensional crystals of the recombinant, full-length channel, as well as crystals in which the C-tail has been completely removed by trypsin digestion. Projection density maps at 7.5 Å resolution closely resemble our previous analysis of the C-terminal truncation mutant (9). A difference map between the full length and trypsin-treated channels suggests that there are small but significant shifts in protein density upon removal of the C-tail.     

A parametric model for estimation of dispersal patterns applied to five passerine spatially structured populations

Natal dispersal capture-recapture data from five fragmented populations of house sparrows, great tits, and blue tits were analyzed using maximum likelihood methods. A new two-parametric distribution was constructed that includes four distributions previously used as special cases in the literature. Dispersal standard deviations were estimated at 22.9 km for the house sparrows and ranged from 0.66 to 4.4 km for the tits. Female great tits and blue tits dispersed consistently further than males. Estimates of the shape parameter of the dispersal distribution ranged from 0.66 to 2.27, indicating strong to moderately leptokurtic dispersal displacements. There were significant effects of density on local immigration rates and a consistent tendency for immigration rates to depend underproportionally on local densities. Potential implications of the shape of the dispersal distribution for the spread of invading organisms were investigated and compared with previous results. It is shown that the wave speed, for a given dispersal standard deviation, depends only to some extent on leptokurtosis, provided that the intrinsic growth rate of the population is moderate or small. In estimating the dispersal standard deviation, however, incorrect assumptions about the degree of leptokurtosis can lead to a large bias in estimation and predictions.     

Towards a unified genetic map for diploid roses

We have constructed the first integrated consensus map (ICM) for rose, based on the information of four diploid populations and more than 1,000 initial markers. The single population maps are linked via 59 bridge markers, on average 8.4 per linkage group (LG). The integrated map comprises 597 markers, 206 of which are sequence-based, distributed over a length of 530?cM on seven LGs. By using a larger effective population size and therefore higher marker density, the marker order in the ICM is more reliable than in the single population maps. This is supported by a more even marker distribution and a decrease in gap sizes in the consensus map as compared to the single population maps. This unified map establishes a standard nomenclature for rose LGs, and presents the location of important ornamental traits, such as self-incompatibility, black spot resistance (Rdr1), scent production and recurrent blooming. In total, the consensus map includes locations for 10 phenotypic single loci, QTLs for 7 different traits and 51 ESTs or gene-based molecular markers. This consensus map combines for the first time the information for traits with high relevance for rose variety development. It will serve as a tool for selective breeding and marker assisted selection. It will benefit future efforts of the rose community to sequence the whole rose genome and will be useful for synteny studies in the Rosaceae family and especially in the section Rosoideae.     

Analysis of the Distribution of Bootstrap Tree Lengths Using the Maximum Parsimony Method

The bootstrap is an important tool for estimating the confidence interval of monophyletic groups within phylogenies. Although bootstrap analyses are used in most evolutionary studies, there is no clear consensus as how best to interpret bootstrap probability values. To study further the bootstrap method, nine small subunit ribosomal DNA (SSU rDNA) data sets were submitted to bootstrapped maximum parsimony (MP) analyses using unweighted and weighted sequence positions. Analyses of the lengths (i.e., parsimony steps) of the bootstrap trees show that the shape and mean of the bootstrap tree distribution may provide important insights into the evolutionary signal within the sequence data. With complex phylogenies containing nodes defined by short internal branches (multifurcations), the mean of the bootstrap tree distribution may differ by 2 standard deviations from the length of the best tree found from the original data set. Weighting sequence positions significantly increases the bootstrap values at internal nodes. There may, however, be strong bootstrap support for conflicting species groupings among different data sets. This phenomenon appears to result from a correlation between the topology of the tree used to create the weights and the topology of the bootstrap consensus tree inferred from the MP analysis of these weighted data. The analyses also show that characteristics of the bootstrap tree distribution (e.g., skewness) may be used to choose between alternative weighting schemes for phylogenetic analyses.     

Bootstrap variance estimators for the parameters of small-sample sensory-performance functions

The bootstrap method, due to Bradley Efron, is a powerful, general method for estimating a variance or standard deviation by repeatedly resampling the given set of experimental data. The method is applied here to the problem of estimating the standard deviation of the estimated midpoint and spread of a sensory-performance function based on data sets comprising 15–25 trials. The performance of the bootstrap estimator was assessed in Monte Carlo studies against another general estimator obtained by the classical combination-of-observations or incremental method. The bootstrap method proved clearly superior to the incremental method, yielding much smaller percentage biases and much greater efficiencies. Its use in the analysis of sensory-performance data may be particularly appropriate when traditional asymptotic procedures, including the probittransformation approach, become unreliable.     

Heterogeneity in Rates of Recombination across the Mouse Genome

If loci are randomly distributed on a physical map, the density of markers on a genetic map will be inversely proportional to recombination rate. First proposed by MARY LYON, we have used this idea to estimate recombination rates from the Drosophila melanogaster linkage map. These results were compared with results of two other studies that estimated regional recombination rates in D. melanogaster using both physical and genetic maps. The three methods were largely concordant in identifying large-scale genomic patterns of recombination. The marker density method was then applied to the Mus musculus microsatellite linkage map. The distribution of microsatellites provided evidence for heterogeneity in recombination rates. Centromeric regions for several mouse chromosomes had significantly greater numbers of markers than expected, suggesting that recombination rates were lower in these regions. In contrast, most telomeric regions contained significantly fewer markers than expected. This indicates that recombination rates are elevated at the telomeres of many mouse chromosomes and is consistent with a comparison of the genetic and cytogenetic maps in these regions. The density of markers on a genetic map may provide a generally useful way to estimate regional recombination rates in species for which genetic, but not physical, maps are available.     

兴安落叶松(Larix gmelinii)林林窗分布规律的小波分析研究

采用小波分析的方法对黑龙江省大兴安岭兴安落叶松林的林窗分布进行分析,研究结果表明：兴安落叶松林样带内计算林窗分布百分率小波变换的最佳尺度为１０ｍ 。林窗分布的疏密变化尺度为２０ｍ 左右,在样地中２０×２０ｍ ２的小区域内具有较为稳定的分布特点。在样带中林窗分布呈斑块状,且斑块分布随样带的海拔的升高呈间断性分布。小波分析被证明是植被空间格局研究的简捷可靠的新方法。     

Genomic analysis in maritime pine (Pinus pinaster). Comparison of two RAPD maps using selfed and open-pollinated seeds of the same individual

Two genomic maps were constructed for one individual tree of maritime pine, Pinus pinaster Ait., using a common set of 263 RAPD markers (random amplified polymorphic DNA). The RAPD markers were chosen from a larger number of polymorphic RAPD fragments on the basis of repeatability and inheritance in a three-generation pedigree. The maps were constructed from two independent mapping samples of 62 megagametophytes (In) from a self cross and from an open-pollinated cross. The markers were grouped (LOD≥4; θ≤0.25) and assigned to 13 major and 5 minor linkage groups. Two framework maps were constructed using the ordering criterion of interval support≥3. Comparison of the two framework maps suggested that the locus order was incorrect for 2% of the framework markers. A bootstrap analysis showed that this error rate was representative for our data set. The results showed that framework maps constructed using RAPD markers were repeatable and that differences in locus order for maps of different genotypes or species could result from chance. The total map distance was 1380 cM, and the map provided coverage of approximately 90% of the genome.     

Estimation of variance in single-particle reconstruction using the bootstrap technique

Density maps of a molecule obtained by single-particle reconstruction from thousands of molecule projections exhibit strong changes in local definition and reproducibility, as a consequence of conformational variability of the molecule and non-stoichiometry of ligand binding. These changes complicate the interpretation of density maps in terms of molecular structure. A three-dimensional (3-D) variance map provides an effective tool to assess the structural definition in each volume element. In this work, the different contributions to the 3-D variance in a single-particle reconstruction are discussed, and an effective method for the estimation of the 3-D variance map is proposed, using a bootstrap technique of sampling. Computations with test data confirm the viability, computational efficiency, and accuracy of the method under conditions encountered in practical circumstances.     

A data mining approach for analyzing density maps representing macromolecular structures

Results of electron microscopy-based three-dimensional reconstructions of macromolecules or their complexes are usually stored as density maps. Each point ("voxel") in the map represents a density value and one approach for studying details of the map is to display an isosurface enclosing areas of interest. We have taken a data mining approach not only focusing on the areas of immediate interest but determining all possible separate entities ("blobs") from a density map. After the entire density map is analyzed with our mining program BLOBBER, properties of all detected blobs can be browsed and sets of blobs can be visualized using our VIZBLOB program. Since BLOBBER analyzes density maps using only density information and relates it to spatial relationships, BLOBBER can be used to analyze symmetrical or asymmetrical density maps from any source. To test our program we have analyzed published bacteriophage PRD1 reconstructions. We identified various structural details ranging from individual proteins to major complexes such as the whole capsid shell and more elaborate details of possible connections between membrane interfaces. This approach can also be a useful preprocessing tool for visualizing reconstructions.     

人类群体遗传空间结构的"克立格"模型

通过将“克立格”技术应用于人类群体遗传学领域,构建了人类群体遗传空间结构的“克立格”模型,并论述了其原理和计算方法。以HLA-A基因座为例,应用“克立格”模型,定量分析了中国人群HLA-A基因座的空间遗传异质性;对HLA-A基因频率的空间数据矩阵进行了主成分分析,进而定义了人类群体遗传结构的综合遗传测度(SPC),绘制了综合遗传测度和主成分(PC)的“克立格”地图,分析了其群体遗传空间结构特性。与其他空间插值或平滑方法相比,人类群体遗传空间结构的“克立格”模型具有明显优点：1)“克立格”估计以空间遗传变异函数模型为基础,在绘制空间遗传结构地图之前,可利用变异函数模型定量分析所研究基因座(或多基因座)的空间遗传异质性;2)“克立格”插值方法是真正意义上的无偏估计模型,它利用待估区域周围的已知群体遗传调查点数据,并充分考虑调查点的空间影响范围,给出待估区域的最优估计值;3)“克立格”模型允许估计插值误差,这种插值误差既可用于评价空间估计效果,又可通过绘制误差地图指导在误差过高的地点增加新的群体遗传调查样本点,以优化估计效果。然而,人类群体遗传空间结构的“克立格”模型也存在一定缺点：1)若不能用任何理论遗传变异函数模型拟合观察遗传变异函数值,则不能建立“克立格”模型;2)若理论遗传变异函数的拟合优度很低,则据此建立的“克立格”模型的估计标准差在整个空间范围内会很大,此时“克立格”模型不适用于估计群体遗传空间结构。出现上述两种情形时,应选用不考虑空间相关性的空间随机插值方法绘制群体遗传结构地图,如基因绘图软件中的Cavalli-Sforza方法,反向距离加权法和条样函数插值法等。     

Projection structure of a N-terminal deletion mutant of connexin 26 channel with decreased central pore density

Gated gap junction channels are important cellular conduits for establishing and maintaining intercellular communication. The three-dimensional structure of a mutant human connexin 26 (Cx26M34A) by electron cryocrystallography revealed a plug-like density in the channel pore suggesting that physical blockage of the pore may be one mechanism of closure (Oshima et al. 2007, Proc Natl Acad Sci USA 104: 10034-10039). However, it remains to be determined what part of the sequence contributes to the plug. Here, we present the projection structure of an N-terminus deletion of Cx26M34A missing amino acids 2 to 7 (Cx26M34Adel2-7) crystallized in the same two-dimensional crystal form. A 10 A resolution projection map of Cx26M34Adel2-7 revealed that the plug density was dramatically reduced in comparison with that found in full-length Cx26 channel. The difference map between the deletion and full-length Cx26M34A channels strongly suggests that the N-terminus of connexin contributes to the plug for the physical closure of gap junction channels.     

A bootstrap test for the analysis of microarray experiments with a very small number of replications

In microarray studies it is common that the number of replications (i.e. the sample size) is small and that the distribution of expression values differs from normality. In this situation, permutation and bootstrap tests may be appropriate for the identification of differentially expressed genes. However, unlike bootstrap tests, permutation tests are not suitable for very small sample sizes, such as three per group. A variety of different bootstrap tests exists. For example, it is possible to adjust the data to have a common mean before the bootstrap samples are drawn. For small significance levels, which can occur when a large number of genes is investigated, the original bootstrap test, as well as a bootstrap test suggested for the Behrens-Fisher problem, have no power in cases of very small sample sizes. In contrast, the modified test based on adjusted data is powerful. Using a Monte Carlo simulation study, we demonstrate that the difference in power can be huge. In addition, the different tests are illustrated using microarray data.     

An integrated genetic linkage map of pepper (Capsicum spp.)

An integrated genetic map of pepper including 6 distinct progenies and consisting of 2262 markers covering 1832 cM was constructed using pooled data from six individual maps by the Keygene proprietary software package INTMAP. The map included: 1528 AFLP, 440 RFLP, 288 RAPD and several known gene sequences, isozymes and morphological markers. In total, 320 anchor markers (common markers in at least two individual maps) were used for map integration. Most anchor markers (265) were common to two maps, while 27, 26 and 5 markers were common to three, four and five maps, respectively. Map integration improved the average marker density in the genome to 1 marker per 0.8 cM compared to 1 marker per 2.1 cM in the most dense individual map. In addition, the number of gaps of at least 10 cM between adjacent markers was reduced in the integrated map. Although marker density and genome coverage were improved in the integrated map, several small linkage groups remained, indicating that further marker saturation will be needed in order to obtain a full coverage of the pepper genome. The integrated map can be used as a reference for future mapping studies in Capsicum and to improve the utilization of molecular markers for pepper breeding.These authors contributed equally to the work described in this paper(e-mail:     

Detection of two-component mixtures of lognormal distributions in grouped, doubly truncated data: analysis of red blood cell volume distributions

We have examined the statistical requirements for the detection of mixtures of two lognormal distributions in doubly truncated data when the sample size is large. The expectation-maximization algorithm was used for parameter estimation. A bootstrap approach was used to test for a mixture of distributions using the likelihood ratio statistic. Analysis of computer simulated mixtures showed that as the ratio of the difference between the means to the minimum standard deviation increases, the power for detection also increases and the accuracy of parameter estimates improves. These procedures were used to examine the distribution of red blood cell volume in blood samples. Each distribution was doubly truncated to eliminate artifactual frequency counts and tested for best fit to a single lognormal distribution or a mixture of two lognormal distributions. A single population was found in samples obtained from 60 healthy individuals. Two subpopulations of cells were detected in 25 of 27 mixtures of blood prepared in vitro. Analyses of mixtures of blood from 40 patients treated for iron-deficiency anemia showed that subpopulations could be detected in all by 6 weeks after onset of treatment. To determine if two-component mixtures could be detected, distributions were examined from untransfused patients with refractory anemia. In two patients with inherited sideroblastic anemia a mixture of microcytic and normocytic cells was found, while in the third patient a single population of microcytic cells was identified. In two family members previously identified as carriers of inherited sideroblastic anemia, mixtures of microcytic and normocytic subpopulations were found. Twenty-five patients with acquired myelodysplastic anemia were examined. A good fit to a mixture of subpopulations containing abnormal microcytic or macrocytic cells was found in two. We have demonstrated that with large sample sizes, mixtures of distributions can be detected even when distributions appear to be unimodal. These statistical techniques provide a means to characterize and quantify alterations in erythrocyte subpopulations in anemia but could also be applied to any set of grouped, doubly truncated data to test for the presence of a mixture of two lognormal distributions.     

Analysing botanical collecting effort in Amazonia and correcting for it in species range estimation

Aim The aim of this study is to analyse the distribution pattern of the botanical collecting effort in Amazonia so that it can be accounted for when interpreting phytogeographical patterns such as inferred species ranges. We also develop a mechanistic and transparent method for taking into account the bias in collecting effort when estimating likelihoods of species occurrences. Location Amazonia, Neotropics. Methods We utilized electronic data sets of georeferenced herbarium collections (1,063,530 in total). We plotted collecting localities (68,246 in total) on maps overlaid with 1° and 0.5° square grids, and analysed collecting effort using a geographical information system (GIS). We also drew a map of Thiessen polygons, using collecting localities as polygon centres, to visualize collecting density in a scale‐independent way. We then created a ‘collecting activity landscape’ in which well‐collected areas appear as peaks and poorly studied areas as valleys. We demonstrate how this surface can be utilized when estimating species distributions. Results The data available to us confirm that botanical collecting activity is still severely biased in Amazonia. The uncollected area represents 43% of the total area of Amazonia, while another 28% is poorly collected and only 2% can be considered relatively well collected. The Thiessen polygon network represents an improvement in the presentation of collecting intensity compared with square grids. Main conclusions The maps of botanical collecting effort in the Neotropics should be used for visually correcting phytogeographical interpretations. With the help of GIS applications the observed spatial bias in collecting effort can be utilized in estimating the likelihood of occurrence of species in a repeatable manner. These estimates, in turn, can be used for various purposes in basic and applied science as well as in decision‐making. The biased collecting effort should, in the long run, be corrected by further field work in unexplored areas, which can be identified with the maps presented here.     

An integrated map of Arabidopsis thaliana for functional analysis of its genome sequence.

The genome of the model plant species Arabidopsis thaliana has recently been sequenced. To accelerate its current genome research, we developed a whole-genome, BAC/BIBAC-based, integrated physical, genetic, and sequence map of the A. thaliana ecotype Columbia. This new map was constructed from the clones of a new plant-transformation-competent BIBAC library and is integrated with the existing sequence map. The clones were restriction fingerprinted by DNA sequencing gel-based electrophoresis, assembled into contigs, and anchored to an existing genetic map. The map consists of 194 BAC/BIBAC contigs, spanning 126 Mb of the 130-Mb Arabidopsis genome. A total of 120 contigs, spanning 114 Mb, were anchored to the chromosomes of Arabidopsis. Accuracy of the integrated map was verified using the existing physical and sequence maps and numerous DNA markers. Integration of the new map with the sequence map has enabled gap closure of the sequence map and will facilitate functional analysis of the genome sequence. The method used here has been demonstrated to be sufficient for whole-genome physical mapping from large-insert random bacterial clones and thus is applicable to rapid development of whole-genome physical maps for other species.