1000 resultados para obtainment construction
Resumo:
We present a systematic, practical approach to developing risk prediction systems, suitable for use with large databases of medical information. An important part of this approach is a novel feature selection algorithm which uses the area under the receiver operating characteristic (ROC) curve to measure the expected discriminative power of different sets of predictor variables. We describe this algorithm and use it to select variables to predict risk of a specific adverse pregnancy outcome: failure to progress in labour. Neural network, logistic regression and hierarchical Bayesian risk prediction models are constructed, all of which achieve close to the limit of performance attainable on this prediction task. We show that better prediction performance requires more discriminative clinical information rather than improved modelling techniques. It is also shown that better diagnostic criteria in clinical records would greatly assist the development of systems to predict risk in pregnancy.
Resumo:
Perhaps the most difficult job of the ecotoxicologist is extrapolating data calculated from laboratory experiments with high precision and accuracy into the real world of highly-dynamics aquatic environments. The establishment of baseline laboratory toxicity testing data for individual compounds and ecologically important and field studies serve as a precursor to ecosystem level studies needed for ecological risk assessment. The first stage in the field portion of risk assessment is the determination of actual environmental concentrations of the contaminant being studied and matching those concentrations with laboratory toxicity tests. Risk estimates can be produced via risk quotients that would determine the probability that adverse effects may occur. In this first stage of risk assessment, environmental realism is often not achieved. This is due, in part, to the fact that single-species laboratory toxicity tests, while highly controlled, do not account for the complex interactions (Chemical, physical, and biological) that take place in the natural environment. By controlling as many variables in the laboratory as possible, an experiment can be produced in such a fashion that real effects from a compound can be determined for a particular test organism. This type of approach obviously makes comparison with real world data most difficult. Conversely, field oriented studies fall short in the interpretation of ecological risk assessment because of low statistical power, lack of adequate replicaiton, and the enormous amount of time and money needed to perform such studies. Unlike a controlled laboratory bioassay, many other stressors other than the chemical compound in question affect organisms in the environment. These stressors range from natural occurrences (such as changes in temperature, salinity, and community interactions) to other confounding anthropogenic inputs. Therefore, an improved aquatic toxicity test that will enhance environmental realism and increase the accuracy of future ecotoxicological risk assessments is needed.
Resumo:
利用聚合酶链式反应(PCR)技术从Alcaligenes eutrophus H16染色体DNA中扩增并克隆了调控聚-3-羟基丁酸(poly-3-hydroxy-butyrate,PHB)生物合成的两个关键酶基因:依赖NADPH的乙酰乙酰CoA还原酶基因(phbB)和PHB合成酶基因(phbC)。限制性内切酶图谱和核苷酸序列分析证实了克隆结果,并表明克隆的基因与国外所报道的有很高的同源性。经过基因拼接,构建了块茎特异性表达的高等植物表达载体pPSAGB(嵌合phbB)、pBIBGC(嵌合phbC)和pPSAGCB(嵌合phbB和phbC)。并以试管薯(microtuber)为外植体经Agrobacterium介导转化了虎头、京丰、Bintje、Favorita、高原4号和88-5共6个马铃薯品种,获得49个株系。经PCR检测导入phbB的株系共有44个,对其中30个株系进行DNA dot blot分析,结果表明phbC导入呈阳性的株系有20个。深入的鉴定工作还在进行中。
Resumo:
聚-β-羟基链烷酸(PHA)是许多微生物作为碳源、能源的一类贮藏性聚酯,具有广泛的应用价值。该聚酯可被微生物完全降解且有与塑料相似的性质,因而研究并提高PHA在植物中的合成为解决环境污染提供了新的解决途径。 聚-β-羟基于酸酯(PHB)是研究的最早、研究的最清楚的一种PHA。用聚合酶链式反应扩增并克隆了真养产碱杆菌(Alcaligenes eutrophus)中合成PHB的一个关键酶——3-酮硫裂解酶基因phbA。DNA序列分析表明所克隆的基因与国外报道序列同源性很高,只有一个碱基对的区别。为了检测该基因的功能及导肽的定位效率,构建了带有导肽基因的组成型表达载体,由根癌农杆菌介导转化烟草(Nicotiana tabacum cv. Wisconsin 38)得到转基因植株。蛋白质电泳结果表明导肽可以将外源蛋白定位于质体,phbA基因能翻译成相应大小的蛋白。酶活性分析证实了转基因烟草中phbA编码的3-酮硫裂解酶可以催化乙酰-CoA合成乙酰乙酰-CoA。 将携有导肽序列的phbC(编码PHB合酶)和phbB(编码乙酰乙酰-CoA还原酶)连入pBIB-HYG得到组成型表达载体pZCB,用冻融法转入根癌农杆菌,介导转化烟草。烟草为已获得的具有卡那霉素抗性整合并表达phbA的转基因烟草。通过二次转化将携有潮霉素抗性的phbB基因和phbC基因导入已整合phbA的烟草,各基因均由质体导肽控制,最后得到整合PHB合成的三个酶基因的转基因烟草。转基因烟草经PCR、PCR-Southern检测,初步确定整合phbB和phbC烟草植株。以气相色谱初步分析,转基因烟草中PHB的含量可达鲜重的0.233%。 结果表明phbB和phbC基因可以在真核表达系统中编码相应的蛋白。通过色素分析、荧光动力学等手段分析了PHB在叶绿体中的累积对其功能的影响。 为了提高底物乙酰-CoA的供应能力及减少惰性聚酯对植物体的伤害,分离了种子特异性启动子和质体导肽序列,利用忆经克隆的合成PHB的三个关键酶基因,通过一系列DNA重组,分别构建了含有种子特异性启动子的嵌合phbC、phbB的二价表达载体pSCB及嵌合phbC、phbA、phbB的三价表达载体pSCAB,并由导肽将基因表达产物定位于质体。经根癌农杆菌介导转化油菜(Brassica napus L.) H165,获得转基因油菜植株,并进行了PCR、Southern blot及RT-PCR-DNA杂交等分检测。结果表明,三基因已经分别整合到相应的转基因油菜中,并已在转录水平表达。同时转化了油菜不育系、恢复系和保持系,获得批量转化株,并移入温室栽培。