家蚕抗菌肽在毕赤酵母中的表达

目的：用毕赤酵母真核系统表达有抑菌活性的家蚕抗菌肽(cecropin-XJ)。将pGEX-4T-1-cecropin-XJ上的抗菌肽基因cecropin—XJ克隆至穿梭质粒pSuperY上,用Bln Ⅰ酶切使之线性化后,采用电击法转化酵母SMD1168,转化子用小瓶发酵,经SDS—PAGE检测,表达产物可以在α信号因子的引导下,分泌到培养基中,且表达产物具有明显抑菌活性。     

Rapid quantification of viable legionellae in water and biofilm using ethidium monoazide coupled with real‐time quantitative PCR

Aims: To optimize ethidium monoazide (EMA) coupled with real‐time quantitative PCR (qPCR) and to evaluate its environmental applicability on quantifying viable legionellae in water and biofilm of cooling towers and hot water systems. Methods and Results: EMA (0·9–45·5 μg ml^?1) and propidium monoazide (PMA, 0·9 and 2·3 μg ml^?1) combined with qPCR (i.e. EMA‐qPCR and PMA‐qPCR, respectively) were applied to unheated and heated (70°C for 30 min) Legionella pneumophila to quantify viable cells, which was also simultaneously determined by BacLight Bacterial Viability kit with epifluorogenic microscopic enumeration (BacLight‐EM). The effects of nontarget microflora and sample matrix on the performance of EMA‐qPCR were also evaluated. In comparison with BacLight‐EM results, qPCR with EMA at 2·3 μg ml^?1 was determined as the optimal EMA‐qPCR assay, which performed equally well as PMA‐qPCR for unheated Leg. pneumophila but better than PMA‐qPCR for heated Leg. pneumophila (P < 0·05). Moreover, qPCR with EMA at 2·3 μg ml^?1 accurately quantified viable Leg. pneumophila, Legionella anisa and Legionella‐like amoebal pathogens 6 (LLAP 6) without interferences by heated legionellae, unheated nonlegionellae cells and cooling tower water matrix (P > 0·05). As for water and biofilm samples collected from cooling towers and hot water systems, the viable legionellae counts determined by EMA‐qPCR were mostly greater than the culturable counts by culture assay but consistently lower than the total cell counts quantified by qPCR. Conclusions: The qPCR with EMA at 2·3 μg ml^?1 may accurately quantify viable legionellae (including fastidious LLAP 6) and Leg. pneumophila pretreated with superheating and is applicable for water and biofilm samples obtained from cooling towers and hot water systems. Significance and Impact of the Study: The EMA‐qPCR assay may be useful in environmental surveillance for viable legionellae and in evaluation of superheating efficacy against legionellae.     

Tet调控STGC3基因表达CNE2细胞系的建立及其功能初步研究

利用Tet-on调控系统,建立受强力霉素诱导STGC3基因表达的CNE2细胞系,为进一步研究STGC3的功能提供了一个理想的实验平台.先后将调控质粒pTet-on和反应质粒pTRE-STGC3转染入CNE2细胞,并用G418和潮霉素分别进行两轮筛选,运用RT-PCR选择对强力霉素诱导敏感的细胞克隆.用不同浓度强力霉素诱导CNE2/Tet/pTRE-STGC3细胞,RT-PCR检测STGC3的表达,确定强力霉素的最佳诱导浓度.采用此浓度的强力霉素分别诱导CNE2、CNE2/Tet/pTRE、CNE2/Tet/pTRE-STGC3三组细胞,测定细胞的生长曲线、克隆形成率和细胞周期分布.诱导STGC3基因高表达,CNE2细胞增殖速度显著减慢(P<0.05),克隆形成能力显著降低(P<0.01);流式细胞仪检测结果显示,瘤细胞群体中处于G0/G1期细胞数增加,细胞阻滞于G0/G1期.Tet调控STGC3基因表达CNE2细胞系的成功建立,为进一步研究STGC3基因的功能提供一个理想的细胞模型.     

GAL4/UAS系统在转基因技术中的应用研究进展

GAL4/UAS系统是一种转基因技术体系,其原理是利用特定的启动子或增强子,以组织特异性的方式激活酵母转录激活子GAL4的表达,GAL4又以同样的方式引起GAL4反应元件(UAS)-靶基因的转录。GAL4/UAS系统的关键点在于:GAL4基因和UAS-靶基因分别存在于两个转基因系中。GAL4转基因系中有转录激活子,但没有靶基因;在UAS-靶基因系中,转录激活子不存在,因而靶基因处于沉默状态,只有将GAL4转基因系与UAS-靶基因系进行杂交,才可能产生表达靶基因的后代。本文综述了GAL4/UAS系统的建立及其研究应用。     

Distributed File System Virtualization Techniques Supporting On-Demand Virtual Machine Environments for Grid Computing

This paper presents a data management solution which allows fast Virtual Machine (VM) instantiation and efficient run-time execution to support VMs as execution environments in Grid computing. It is based on novel distributed file system virtualization techniques and is unique in that: (1) it provides on-demand cross-domain access to VM state for unmodified VM monitors; (2) it enables private file system channels for VM instantiation by secure tunneling and session-key based authentication; (3) it supports user-level and write-back disk caches, per-application caching policies and middleware-driven consistency models; and (4) it leverages application-specific meta-data associated with files to expedite data transfers. The paper reports on its performance in wide-area setups using VMware-based VMs. Results show that the solution delivers performance over 30% better than native NFS and with warm caches it can bring the application-perceived overheads below 10% compared to a local-disk setup. The solution also allows a VM with 1.6 GB virtual disk and 320 MB virtual memory to be cloned within 160 seconds for the first clone and within 25 seconds for subsequent clones. Ming Zhao is a PhD candidate in the department of Electrical and Computer Engineering and a member of the Advance Computing and Information Systems Laboratory, at University of Florida. He received the degrees of BE and ME from Tsinghua University. His research interests are in the areas of computer architecture, operating systems and distributed computing. Jian Zhang is a PhD student in the Department of Electrical and Computer Engineering at University of Florida and a member of the Advance Computing and Information Systems Laboratory (ACIS). Her research interest is in virtual machines and Grid computing. She is a member of the IEEE and the ACM. Renato J. Figueiredo received the B.S. and M.S. degrees in Electrical Engineering from the Universidade de Campinas in 1994 and 1995, respectively, and the Ph.D. degree in Electrical and Computer Engineering from Purdue University in 2001. From 2001 until 2002 he was on the faculty of the School of Electrical and Computer Engineering of Northwestern University at Evanston, Illinois. In 2002 he joined the Department of Electrical and Computer Engineering of the University of Florida as an Assistant Professor. His research interests are in the areas of computer architecture, operating systems, and distributed systems.     

The Hilltopping Mating System of Leschenaultia adusta (Loew) (Diptera: Tachinidae)

Males of the tachinid fly Leschenaultia adusta perch on small trees and shrubs on the highest parts of Usery Peak in central Arizona. Individuals select twig perches on the downwind side of these plants and fly out spontaneously from time to time or in response to another passing insect. Conspecific males elicit chases that on occasion escalate into elaborate, high-speed pursuit flights that go back and forth near the plant for several minutes. Although several males sometimes perch together briefly in the same plant, typically only one individual remains at a site for more than an hour on any given day. These site-faithful males can be considered territorial residents; they constituted about one-quarter of the males marked during the study. More than half of these residents returned to the same perch plant for two or more days. Perch plants varied in their attractiveness to male flies; male preferences were largely consistent across two years of study. Given that females were occasionally observed mating at male-occupied plants, we place the mating system of L. adusta within the hilltopping territorial category in which males compete for landmark perching sites attractive to receptive females. As is true for other hilltopping insects, receptive females of L. adusta appear to be rare and widely distributed.     

Assessment of the potential of Napier grass (Pennisetum purpureum) varieties as trap plants for management of Chilo partellus

Ten Napier grass [Pennisetum purpureum Schumach (Poaceae)] varieties, used in various parts of Kenya as animal fodder, were tested for their potential role as a trap crop in the management of the gramineous spotted stemborer, Chilo partellus (Swinhoe) (Lepidoptera: Crambidae), through habitat manipulation. Oviposition preference and larval survival and development were determined for each of these varieties under laboratory and screen‐house conditions. Two‐choice tests revealed that seven of the varieties tested were preferentially chosen by gravid female moths for oviposition over a susceptible maize cv. Inbred A. Larval survival was significantly lower on the majority of the Napier grass varieties (about 10%) than on maize cv. Inbred A (about 60%). Similarly, larval development was about 2 weeks longer on the majority of the Napier grass varieties. It was concluded that four of the varieties tested (Bana, Ex‐Machakos, Gold Coast, and Ex‐Nyanza‐2) had potential for use as trap crops in C. partellus management because they were more preferred by the borer moths for oviposition and allowed minimal survival of the larvae. This minimal larval survival is desirable in the conservation of C. partellus natural enemies when the cultivated host plant is out of season.     

Why be a rewarding trap? The evolution of floral rewards in Arum (Araceae), a genus characterized by saprophilous pollination systems

Plant species of the genus Arum typically have lure-and-trap pollination systems that are saprophilous (i.e. attracting flies or beetles searching for breeding sites in decaying organic matter). They have been assumed to always attract and trap their pollinators by deception because the inflorescences provide unsuitable breeding grounds for pollinators. The present study explored the possibility that one species, Arum creticum Boiss. & Heldr., which has yellow, sweet-smelling inflorescences, rewards its pollinators and that this increases its success in attracting pollinators over its close relative, Arum idaeum Coust. & Gadoger. The relationship between rewards provided, floral structure, insect attraction, and pollen import and export was examined in two naturally occurring sympatric populations of A. creticum , A. idaeum , and their natural hybrids. The results showed that plants providing more pollen were visited by larger numbers of females of a mining bee Lasioglossum marginatum Brullé as well as adults and nymphs of a hemipteran bug Dionconotus cruentatus creticus Heiss. In A. creticum , L. marginatum was found to be a better outcrossing vector than D. cruentatus in areas where both pollinators occurred because L. marginatum individuals carried more pollen on their bodies and travelled greater distances between inflorescences, thus increasing the potential for outcrossing. The finding that floral rewards may result in increased fitness, compared to nonrewarding systems that rely on attracting saprophilic pollinators, suggests that it is possible for rewarding species to evolve from saprophilous systems.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 257–268.     

Designing a conservation plan for protecting the habitat for giant pandas in the Qionglai mountain range, China

Population viability of the giant panda ( Ailuropoda melanoleuca ) is threatened by small population sizes in scattered isolated habitat areas. Designing a conservation plan for protecting and connecting the fragmented habitat will improve the chances for survival of this endangered species. For such a plan, this study assessed the overall habitat suitability for the species in the Qionglai mountain range (Sichuan, China) using Landsat TM imagery acquired in 2001, geographical data, field surveys, and information acquired in previous researches. Results show that the habitat is separated by roads and rivers, as well as by human settlements and cropland areas, into four main habitat blocks. Overlapping these four habitat blocks with the current nature reserve network reveals that only 36% of the total habitat is protected within nature reserves. Thus, the current nature reserve network is failing to preserve essential habitat for dispersal and genetic exchange. In this study, five key areas and four linkage areas were identified and suggested as nature reserves and/or corridors. These areas, together with the six currently established nature reserves in the mountain range, will form a conservation unit for facilitating the exchange of giant panda individuals among previously isolated habitat blocks. Policies recently implemented by the Chinese government, including the Natural Forest Conservation Program (NFCP) and the Grain-to-Green Program (GTGP), could aid in the formation of such a conservation unit.     

Determinants of regional species richness: an empirical analysis of the number of hawkmoth species (Lepidoptera: Sphingidae) on the Malesian archipelago

Aims Major patterns and determinants of the species richness of Sphingidae in the Malesian archipelago were investigated, including a distinction of richness patterns between subfamilies and range‐size classes. Location Southeast Asia, Malesia. Methods Using a compilation of specimen‐label data bases, geographic information system (GIS)‐supported estimates of distributional ranges for all Sphingidae species of Southeast Asia were used to assess the species richness of islands. Range maps for all species and checklists for 114 islands can be found at http://www.sphingidae‐sea.biozentrum.uni‐wuerzburg.de . Potential determinants of the species richness of islands were tested with general linear models. Results The estimated species richness of islands in the region is determined by biogeographical association, seasonality, availability of rain forest and island size. Species–area relationships are linear on a semi‐logarithmic representation, but not on a double‐logarithmic scale. Species richness of all sphingid subfamilies is influenced by biogeography. The presence of large rain‐forest areas affects mainly Smerinthinae, whereas distance from continental Asia is conspicuously irrelevant for this group. Widespread rather than geographically restricted species shape the overall distribution patterns of species richness. The altitudinal range of islands does not significantly affect species‐richness patterns, but its potential effects on geographically restricted species are discussed. Main conclusions As well as being affected by climatic and vegetation parameters, sphingid species richness is strongly influenced by a historical, directional dispersal process from continental Southeast Asia to the Pacific islands. This process did not apply equally to species of different taxonomic groups or range sizes. Widespread species decline in species richness towards the south‐east, whereas geographically restricted species exhibit an inverse pattern of species richness, probably because speciation becomes more important in this group within the more isolated island groups.