Genetic selection for a highly functional cysteine-less membrane protein using site saturation mutagenesis

We describe an efficient method for generating highly functional membrane proteins with variant amino acids at defined positions that couples a modified site saturation strategy with functional genetic selection. We applied this method to the production of a cysteine-less variant of the Crithidia fasciculata inosine-guanosine permease CfNT2 to facilitate biochemical studies using thiol-specific modifying reagents. Of 10 endogenous cysteine residues in CfNT2, two cannot be replaced with serine or alanine without loss of function. High-quality single- and double-mutant libraries were produced by combining a previously reported site saturation mutagenesis scheme based on the Stratagene Quikchange method with a novel gel purification step that effectively eliminated template DNA from the products. Following selection for functional complementation in Saccharomyces cerevisiae cells auxotrophic for purines, several highly functional noncysteine substitutions were efficiently identified at each desired position, allowing the construction of cysteine-less variants of CfNT2 that retained wild-type affinity for inosine. This combination of an improved site saturation mutagenesis technique and positive genetic selection provides a simple and efficient means to identify functional and perhaps unexpected amino acid variants at a desired position.     

马占相思树干液流特征及其与环境因子的关系

运用Granier热消散式探针法对华南丘陵退化荒坡植被恢复先锋树种马占相思(Acaciamangium)树干液流密度进行长期连续观测,并对其周围环境因子如空气温度、空气相对湿度、土壤相对湿度、光合有效辐射和总辐射进行同步观测。通过分析发现马占相思边材厚度与胸径存在显著线性相关关系;马占相思树干液流密度最大值与边材面积具显著相关关系;马占相思树干东、南、西、北4个方位测得的液流密度具显著差异,且各方位相互之间均有显著相关关系;马占相思树干液流每天到达峰值与光合有效辐射和水蒸气压亏缺到达峰值存在一定的时滞,这两个时滞与树高无关,个体间时滞差异在湿季较小,干季较大;干湿季液流平均值和最大值具显著差异,湿季蒸腾水量大于相同时间内干季蒸腾水量;液流的变化与空气温度、空气相对湿度、光合有效辐射、总辐射、水蒸气压亏缺等环境因子的变化具显著相关关系,按相关程度排序为:光合有效辐射>总辐射>水蒸气压亏缺>空气相对湿度>空气温度。     

The extension reached by the minimization of the polarity distances during the evolution of the genetic code

Summary The level reached by the optimization of the polarity distances during the evolution of the genetic code was investigated. The results, although not conclusive, indicate that this optimization level is higher than the data reported in the literature. The results seem compatible with the reaching of an evolutionary minimum, with respect to the optimization of the polarity distances, by the genetic code during its formation.     

基于多目标灰色局势决策的三峡库区防护林类型空间优化配置

基于2007年Landsat TM遥感影像和影响防护林的主导环境因子,对三峡库区的森林立地进行分类,并通过选取水源涵养量、生物量和林分生产力3个指标,利用多目标灰色局势决策模型对库区现有的针叶林、阔叶林、针阔混交林和灌木林4种防护林类型进行空间优化配置.结果表明： 2007年,三峡库区森林立地可划分为40种类型;空间配置优化后,研究区针叶林、阔叶林、针阔混交林和灌木林的面积比例分别为32.55%、29.43%、34.95%和3.07%.与优化前相比,优化后针叶林和灌木林的面积比例分别减少了8.79%和28.55%,阔叶林和针阔混交林分别增加了10.23%和27.11%.通过防护林类型的空间优化,三峡库区整体的水源涵养能力、生物量和林分生产力分别增加14.09×10⁸ m³、0.35×10⁸ t和1.08×10⁶ t.     

土壤微生物量测定方法概述

土壤微生物量是土壤生态系统研究的重要参数之一。常用的土壤微生物量的测定方法主要包括:直接镜检法、熏蒸系列方法、底物诱导系列方法、成分分析法和比色法。对这些具体测定方法、原理及其优缺点进行了简要的评述,并指出了应用这些方法须注意的问题和今后的研究方向。     

一种采集稻纵卷叶螟卵的高效简便新方法

研究了稻纵卷叶螟Cnaphalocrocis medinalis Guenee的"烧杯+湿纱布"卵收集方法,并与"塑料袋收集法"和"塑料杯收集法"进行了比较。结果表明,"烧杯+湿纱布"卵收集法能显著地延长了稻纵卷叶螟成蛾的寿命、提高单雌产卵量和卵孵化率。采用此法收集稻纵卷叶螟的卵更高效和简便。     

Stability of Prussian Blue in Soils of a Former Manufactured Gas Plant Site

Soil contamination with iron-cyanide complexes is a common problem at former manufactured gas plant (MGP) sites. Dissolution of the cyanide, from Prussian Blue (ferric ferrocyanide), creates an environmental hazard, whereas the risk of groundwater contamination depends on the stability of dissolved iron–cyanide complexes. Lack of a standard leaching method to determine the water-soluble (plant-available) cyanide fraction generates potential limitations for implementing remediation strategies like phytoremediation. Applicability of neutral solution extraction to determine the water-soluble cyanide fraction and the stability of Prussian Blue in surface and near-surface soils of an MGP site in Cottbus, undersaturated and unsaturated water conditions, was studied in column leaching and batch extraction experiments. MGP soils used in the long-term tests varied according to the pH (5.0–7.7) and the total cyanide content (40–1718 mg kg^?1). Column leaching, after four months of percolation, still yielded effluent concentrations exceeding the German drinking water limit (> 50 μg L^?1) and the solubility of Prussian Blue reported in the literature (< 1 mg L^?1) from both alkaline and acidic soils. Long-term (1344 h) extraction of MGP soils with distilled water was sufficient to dissolve 97% of the total cyanide from the slightly alkaline soils and up to 78% from the acidic soils. Both experiments revealed that dissolution of ferric ferrocyanide under circum-neutral pH and oxic water conditions is a function of time, where the released amount is dependent on the soil pH and total cyanide content. Unexpectedly high and continuous solubility of Prussian Blue, both in acidic and slightly alkaline MGP soils, implies the need to introduce an additional cyanide fraction (“readily soluble fraction”) to improve and specify cyanide leaching methods. Long-term extraction of cyanide-contaminated soil in neutral solution seems to be a promising approach to evaluate the potential hazard of groundwater pollution at the MGP sites.     

武夷山杉木林凋落物动态初探

用起伏型时间序列法对武夷山国家级自然保护区杉木林凋落物月动态进行模型,结果令人满意,说明起伏型时间序列分析可应用于森林凋落物动态模拟。     

几种红树植物幼苗中可溶性糖的含量

用Ｓｏｍｏｇｙｉ法测定了红海榄、木榄和秋茄３种红树植物幼苗的根、茎、叶和胚轴中可溶性糖的含量。同时．进行了加样回收率试验,回收率平均为１００．４８％,变异系数为２．１４％     

Confidence intervals for the mean of diagnostic test charge data containing zeros

In this paper, we consider the problem of interval estimation for the mean of diagnostic test charges. Diagnostic test charge data may contain zero values, and the nonzero values can often be modeled by a log-normal distribution. Under such a model, we propose three different interval estimation procedures: a percentile-t bootstrap interval based on sufficient statistics and two likelihood-based confidence intervals. For theoretical properties, we show that the two likelihood-based one-sided confidence intervals are only first-order accurate and that the bootstrap-based one-sided confidence interval is second-order accurate. For two-sided confidence intervals, all three proposed methods are second-order accurate. A simulation study in finite-sample sizes suggests all three proposed intervals outperform a widely used minimum variance unbiased estimator (MVUE)-based interval except for the case of one-sided lower end-point intervals when the skewness is very small. Among the proposed one-sided intervals, the bootstrap interval has the best coverage accuracy. For the two-sided intervals, when the sample size is small, the bootstrap method still yields the best coverage accuracy unless the skewness is very small, in which case the bias-corrected ML method has the best accuracy. When the sample size is large, all three proposed intervals have similar coverage accuracy. Finally, we analyze with the proposed methods one real example assessing diagnostic test charges among older adults with depression.