极端耐热纤维素酶Cel74的表达、纯化与特性

从海栖热袍菌中克隆出编码热稳定性的纤维素酶基因,以热激载体pHsh为表达质粒,构建重组质粒phsh—Ceff4,并转化至大肠杆菌中进行表达。基因表达产物通过热处理和离子交换层析,重组酶纯度达电泳纯。对纯化的重组酶酶学性质研究表明,最适反应温度85℃,最适反应pH4．6,pH4．5—6．0之间酶的相对酶活在80％以上。Co^2＋对酶活性有促进作用,Ca^2＋、Mg^2＋、Zn^2＋不影响酶活性,而Cu^2＋、Ni^2＋、Mn^2＋对酶活性有抑制作用。     

4种生态因子对虎斑乌贼幼体生长与存活的影响

旨在研究温度、盐度、饵料种类、投饵量对虎斑乌贼（Sepia pharaonis）幼体生长与存活率的影响,以确定其生长发育的最佳生态条件,为人工育苗提供理论依据。在室内控制条件下,采用单因子试验研究了不同温度（18、21、24、27、30和33℃）、不同盐度（18、21、24、27、30和33）、不同饵料种类（虾糜、活糠虾、卤虫无节幼体、桡足类、死糠虾、虾糜＋强化卤虫后无节幼体、强化卤虫后无节幼体）、活糠虾不同投饵量（0、2、4、6、8和10 g/d）对虎斑乌贼幼体生长发育的影响。结果显示：不同温度对虎斑乌贼幼体生长影响显著（P〈0.05）,最适温度为24-27℃,在最适温度下,存活率为84.4%-91.1%,特定生长率为4.82%-6.13%,存活率（y）与温度（x）的函数关系为y=-0.15x3＋30.637x-447.002（r2=0.923）。幼体适宜盐度为24-33,最适盐度为27,在最适盐度条件下,其存活率为（90.0±5.29）%,特定生长率为（3.71±0.34）%。投喂7种开口饵料,以活糠虾效果最佳,存活率为（96.7±2.94）%,特定生长率为（3.77±0.23）%;强化卤虫后无节幼体效果次之,存活率为（95.6±2.31）%,特定生长率为（2.54±0.15）%,其余各组培养效果均不理想。投喂活糠虾,随着投饵量的增加,其存活率随之提高,摄食量（y）与个体重（x）的回归方程为y=0.227x-0.063（r2=0.921）。     

生态学的时空特性(英文)

 众所周知,几乎所有的生态学特征和现象都受限于一定的时间和空间。因此,相应的科学假设和相关的生态学结论也只能基于这些特定的时空尺度范围。我们利用颇为熟知的事例,引用生态学文献中的具体实例,提纲挈领地论述了时空在生态学研究中的重要性。这些实例包括我们在长白山对云、冷杉(Picea jezoensis, Abies nephrolepis)林林冠结构的模拟、在北美应用遥感和气象方法对碳通量的估算,以及测定湿地生态系统对加温的反应等。文中所涉及的所有生态学现象,对时间和空间都有强烈的依赖性。因而, 从生态学问题的提出,到科学假设的演绎,以至试验设计和综合数据分析,都必须以时、空为前提,才不至于导致荒谬结论。     

Variation in heat shock proteins within tropical and desert species of poeciliid fishes

The 70-kilodalton heat shock protein (hsp70) family of molecular chaperones, which contains both stress-inducible and normally abundant constitutive members, is highly conserved across distantly related taxa. Analysis of this protein family in individuals from an outbred population of tropical topminnows, Poeciliopsis gracilis, showed that while constitutive hsp70 family members showed no variation in protein isoforms, inducibly synthesized hsp70 was polymorphic. Several species of Poeciliopsis adapted to desert environments exhibited lower levels of inducible hsp70 polymorphism than the tropical species, but constitutive forms were identical to those in P. gracilis, as they were in the confamilial species Gambusia affinis. These differences suggest that inducible and constitutive members of this family are under different evolutionary constraints and may indicate differences in their function within the cell. Also, northern desert species of Poeciliopsis synthesize a subset of the inducible hsp70 isoforms seen in tropical species. This distribution supports the theory that ancestral tropical fish migrated northward and colonized desert streams; the subsequent decrease in variation of inducible hsp70 may have been due to genetic drift or a consequence of adaptation to the desert environment. Higher levels of variability were found when the 30- kilodalton heat shock protein (hsp30) family was analyzed within different strains of two desert species of Poeciliopsis and also in wild-caught individuals of Gambusia affinis. In both cases the distribution of hsp30 isoform diversity was similar to that seen previously with allozyme polymorphisms.      

Ethylene-induced Tropism of Trifolium fragiferum L. Stolons

The hypothesis that ethylene regulates prostrate stem growth was investigated using stolons of strawberry clover (Trifolium fragiferum L. var. Salina). Stolons became erect when treated with ethylene or 2-chloroethylphosphonic acid. Curvature was visibly detectable 2 hours after ethylene treatment, and subsequent stem elongation was rapid. Indoleacetic acid application to prostrate stolons caused only a small transitory curvature persisting less than 48 hours. Indoleacetic acid-stimulated curvature was accompanied by an increase in ethylene evolution. When stolon curvature was induced by placing strawberry clover plants in darkness or by applying gibberellic acid, ethylene production did not parallel stolon curvature.     

MicroRNA与肺癌发生、诊断及治疗

微小RNA(miRNAs)是一大类小的非编码RNA,它通过与靶mRNA 3′非翻译区部分互补配对来调节特定基因的表达。近来研究表明,miRNA可作为癌基因或抑癌基因在肺癌发生发展过程中起重要作用。比较癌组织和非癌组织中miRNA表达谱的差异可筛选出部分miRNA分子作为肺癌诊断和预后判断的潜在生物标记。调节具有致癌或抑癌功能的miRNA表达可能成为肺癌治疗新方法,而结合传统放化疗及其敏感性miRNA标志也为肺癌治疗研究提供了新的策略。该文对miRNA在肺癌发生与发展、基因诊断和治疗中的作用做一综述。     

Are generalist Aphidiinae (Hym. Braconidae) mostly cryptic species complexes?

Aphidiinae are mostly composed of specialist parasitoids and the few species described as generalist are suspected to be composed of cryptic specialists, almost indistinguishable based on morphological characteristics. The use of molecular markers has proven to be a useful tool for revealing cryptic species complexes and here we use seven mitochondrial and nuclear gene fragments to study possible genetic differentiation among seven Aphidiinae generalists. Maximum likelihood (ML) trees and Bayesian Poisson tree processes (bPTP) models were conducted on each gene separately and on the seven genes together. The standard cytochrome c oxidase I barcode region appeared to be the most polymorphic and probably the best marker to reveal putative cryptic species. However, we showed with ML trees and bPTP models that a complementary use of mitochondrial and nuclear genes was the most relevant approach to reliably identify cryptic genetic clades in the Aphidiinae. Overall, most of the analysed generalist morphospecies were shown to be composed of subgroups related to the aphid host, some of them revealed as cryptic species by the species delimitation analysis. Further studies are needed to reveal the generality of this result in Aphidiinae.     

荧光标记复合PCR扩增微卫星位点的构建与优化

本研究采用毛细管电泳技术,构建并优化了荧光标记复合PCR同时扩增多个微卫星位点。主要过程为：首先根据设计所扩增微卫星位点的期望长度,将9个微卫星位点分成两组,5个位点用FAM（蓝色）标记,4个位点用HEX（绿色）标记;两种荧光类型分组优化,用琼脂糖胶电泳检测。其次,荧光标记的复合PCR扩增8个中华绒螯蟹样品的9个微卫星位点,采用ABI3730xl毛细管电泳检测,以ROX500（红色）为长度标准物,结果经Genemapper3.5软件 分析,检测结果表明毛细管电泳检测荧光标记复合PCR产物不仅精确读取微卫星位点的长度（分辨率高达1bp）,还能区分微卫星位点复制时滑链所引起的“回声斑”;调整各微卫星位点引物比列使所有位点扩增强弱均匀。最后,逐一检测复合PCR基本参数（dNTP浓度、 PCR程序和模版DNA用量）对复合PCR产物的影响,优化PCR。结果表明通过毛细管电泳检测荧光标记复合PCR产物来读取微卫星位点的基因型具有精确性、高效性和稳定性。